Compositions for pulmonary delivery of long-acting muscarinic antagonists and associated methods and systems

ABSTRACT

Compositions, methods and systems are provided for pulmonary delivery of long-acting muscarinic antagonists and long-acting β 2  adrenergic receptor agonists via a metered dose inhaler. In particular embodiments, the compositions include a suspension medium, active agent particles, and suspending particles, in which the active agent particles and suspending particles form a co-suspension within the suspension medium.

This application claims the benefit of priority of U.S. ProvisionalApplication No. 61/182,565, filed May 29, 2009; U.S. ProvisionalApplication No. 61/258,172, filed Nov. 4, 2009; U.S. ProvisionalApplication No. 61/309,365, filed Mar. 1, 2010; and U.S. ProvisionalApplication No. 61/345,536 filed May 17, 2010.

TECHNICAL FIELD

The present disclosure relates generally to pharmaceutical formulationsand methods for delivery of active agents via the respiratory tract. Incertain aspects, the present disclosure relates to compositions,methods, and systems for pulmonary delivery of long-acting muscarinicantagonists and long-acting β₂ adrenergic receptor agonists via ametered dose inhaler.

BACKGROUND

Methods of targeted drug delivery that deliver an active agent at thesite of action are often desirable. For example, targeted delivery ofactive agents can reduce undesirable side effects, lower dosingrequirements and decrease therapeutic costs. In the context ofrespiratory delivery, inhalers are well known devices for administeringan active agent to a subject's respiratory tract, and several differentinhaler systems are currently commercially available. Three commoninhaler systems include dry powder inhalers, nebulizers and metered doseinhalers (MDIs).

MDIs may be used to deliver medicaments in a solubilized form or as asuspension. Typically, MDIs use a relatively high vapor pressurepropellant to expel aerosolized droplets containing an active agent intothe respiratory tract when the MDI is activated. Dry powder inhalersgenerally rely on the patient's inspiratory efforts to introduce amedicament in a dry powder form to the respiratory tract. On the otherhand, nebulizers form a medicament aerosol to be inhaled by impartingenergy to a liquid solution or suspension.

MDIs are active delivery devices that utilize the pressure generated bya propellant. Conventionally, chlorofluorocarbons (CFCs) have been usedas propellants in MDI systems because of their low toxicity, desirablevapor pressure and suitability for formulation of stable suspensions.However, traditional CFC propellants are understood to have a negativeenvironmental impact, which has led to the development of alternativepropellants that are believed to be more environmentally-friendly, suchas perfluorinated compounds (PFCs) and hydrofluoroalkanes (HFAs).

The active agent to be delivered by a suspension MDI is typicallyprovided as a fine particulate dispersed within a propellant orcombination of two or more propellants (i.e., a propellant “system”). Inorder to form the fine particulates, the active agent is typicallymicronized. Fine particles of active agent suspended in a propellant orpropellant system tend to aggregate or flocculate rapidly. This isparticularly true of active agents present in micronized form. In turn,aggregation or flocculation of these fine particles may complicate thedelivery of the active agent. For example, aggregation or flocculationcan lead to mechanical failures, such as those that might be caused byobstruction of the valve orifice of the aerosol container. Unwantedaggregation or flocculation of drug particles may also lead to rapidsedimentation or creaming of drug particles, and such behavior mayresult in inconsistent dose delivery, which can be particularlytroublesome with highly potent, low dose medicaments. Another problemassociated with such suspension MDI formulations relates to crystalgrowth of the drug during storage, resulting in a decrease over time ofaerosol properties and delivered dose uniformity of such MDIs. Morerecently, solution approaches, such as those disclosed in U.S. Pat. No.6,964,759, have been proposed for MDI formulations containinganticholinergics.

One approach to improve aerosol performance in dry powder inhalers hasbeen to incorporate fine particle carrier particles, such as lactose.Use of such fine excipients has not been investigated to any greatextent for MDIs. A recent report by Young et al., “The influence ofmicronized particulates on the aerosolization properties of pressurizedmetered dose inhalers”; Aerosol Science 40, pgs. 324-337 (2009),suggests that the use of such fine particle carriers in MDIs actuallyresult in a decrease in aerosol performance.

In traditional CFC systems, when the active agent present in an MDIformulation is suspended in the propellant or propellant system,surfactants are often used to coat the surfaces of the active agent inorder to minimize or prevent the problem of aggregation and maintain asubstantially uniform dispersion. The use of surfactants in this manneris sometimes referred to as “stabilizing” the suspension. However, manysurfactants that are soluble and thus effective in CFC systems are noteffective in HFA and PFC propellant systems because such surfactantsexhibit different solubility characteristics in non-CFC propellants.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph, which depicts the particle size distributionexhibited by an exemplary co-suspension composition according to thepresent description, which included glycopyrrolate, a long-actingmuscarinic antagonist, as the active agent. Co-suspension MDIs weresubjected to temperature cycling conditions (alternating 6 h hold timeat −5 or 40° C.) for 12 weeks.

FIG. 2 is a graph, which depicts the particle size distributionexhibited by an exemplary co-suspension composition according to thepresent description, which included glycopyrrolate, a long-actingmuscarinic antagonist, as the active agent. Co-suspension MDIs weresubjected to temperature cycling conditions (alternating 6 h hold timeat −5 or 40° C.) for 24 weeks.

FIG. 3 provides a micrograph illustrating the morphologies of a varietyof suspending particles prepared according to Example 5.

FIG. 4 is a photograph of two vials that allows visualization of aco-suspension formed using active agent particles formed usingglycopyrrolate and suspending particles formed using a saccharide.

FIG. 5 is a graph, which depicts the serum glycopyrrolate concentrationlevel achieved over a period of 24 hours after a single administrationof four different doses of glycopyrrolate delivered from a co-suspensioncomposition as described herein.

FIG. 6 is a graph, which depicts the mean change in FEV₁ from baseline(in liters) experienced in patients over a period of 24 hours afterreceiving a single administration of the indicated dose ofglycopyrrolate formulated in a co-suspension as described herein. Inthis study, Spiriva (18 μg Tiotropium) was included as an activecontrol, and the mean change in FEV₁ from baseline (in liters)experienced in patients receiving a single administration of Spiriva isalso depicted.

FIG. 7 is a bar graph, which depicts the peak change in FEV₁ frombaseline (in liters) experienced in patients after receiving a singleadministration of the indicated dose of glycopyrrolate formulated in aco-suspension as described herein relative to placebo, the area underthe curve of the FEV₁ over 12 hours after dosing, and the area under thecurve of the FEV₁ over 24 hours after dosing relative to placebo acrossthe four doses evaluated. In this study, Spiriva (18 μg Tiotropium) wasincluded as an active control and the results following singleadministration of Spiriva for the above parameters are also depicted inthis figure.

FIG. 8 is a graph, which depicts the proportion of patients whichachieved a greater than 12% change in FEV₁ from baseline and animprovement of 150 mL change from baseline or an absolute improvement of200 mL from baseline regardless of % change in FEV₁ from baseline, afterreceiving a single administration of the indicated doses of aglycopyrrolate co-suspension as described herein. In this study, Spiriva(18 μg Tiotropium) was included as an active control and the resultsfollowing single administration of Spiriva for the above parameter arealso depicted on this figure.

FIG. 9 is a bar graph, which depicts the peak change in inspiratorycapacity experienced in patients after receiving a single administrationof the indicated doses of a glycopyrrolate co-suspension as describedherein. In this study, Spiriva (18 μg Tiotropium) was included as anactive control and the results following single administration ofSpiriva for the above parameter are also depicted on this figure.

FIG. 10 is a bar graph providing the change in FEV₁ AUC achieved inpatients after receiving a single administration of the indicated dosesof a glycopyrrolate co-suspension as described herein. The resultsachieved by the glycopyrrolate co-suspension according to the presentdescription are shown in comparison with the change in FEV₁ AUC reportedin a published study in patients who received a powder formulation ofglycopyrrolate not prepared according to the teachings provided herein.

FIG. 11 is a graph, which depicts the particle size distribution of anexemplary glycopyrrolate co-suspension prepared according to the presentdescription, containing 4.5 μg/actuation delivered dose ofglycopyrrolate and 6 mg/mL suspending particles and subjected totemperature cycling conditions (alternating 6 h hold time at −5 or 40°C.).

FIG. 12 is a graph, which depicts the particle size distribution of anexemplary glycopyrrolate co-suspension prepared according to the presentdescription, containing 36 μg/actuation delivered dose of glycopyrrolateand 6 mg/mL suspending particles and subjected to temperature cyclingconditions (alternating 6 h hold time at −5 or 40° C.).

FIG. 13 is a graph, which depicts the delivered dose through canisterlife of an exemplary glycopyrrolate co-suspension prepared according tothe present description, containing 4.5 μg/actuation delivered dose ofglycopyrrolate and 6 mg/mL suspending particles.

FIG. 14 is a graph, which depicts the delivered dose through canisterlife of an exemplary glycopyrrolate co-suspension prepared according tothe present description, containing 36 μg/actuation delivered dose ofglycopyrrolate and 6 mg/mL suspending particles.

FIG. 15 is a graph, which depicts the particle size distribution of anexemplary glycopyrrolate co-suspension prepared according to the presentdescription, containing 36 μg/actuation delivered dose of glycopyrrolateand 6 mg/mL suspending particles and subjected to 12 months storage at25° C./60% RH unprotected.

FIG. 16 is a graph, which depicts the mean delivered dose throughcanister life of an exemplary glycopyrrolate co-suspension preparedaccording to the present description, containing 32 μg/actuationdelivered dose of glycopyrrolate and 6 mg/mL suspending particles andsubjected to temperature cycling conditions (alternating 6 h hold timeat −5 or 40° C.).

FIG. 17 is a graph, which depicts the particle size distribution of anexemplary glycopyrrolate co-suspension prepared according to the presentdescription, containing 32 μg/actuation delivered dose of glycopyrrolateand 6 mg/mL suspending particles and subjected to temperature cyclingconditions (alternating 6 h hold time at −5 or 40° C.)

FIG. 18 is a graph, which depicts the particle size distribution of anexemplary glycopyrrolate co-suspension prepared according to the presentdescription, containing 24 μg/actuation delivered dose of glycopyrrolateand 6 mg/mL suspending particles and subjected to 6 weeks storage at 50°C./ambient relative humidity and 12 weeks at 40° C.

FIG. 19 is a photograph that allows visualization of co-suspensioncompositions prepared according to the present description which includeformoterol fumarate active agent particles.

FIG. 20 is a graph, which depicts the delivered dose uniformity achievedby formoterol fumarate co-suspension compositions prepared according tothe present description.

FIG. 21 is a graph, which depicts the aerodynamic particle sizedistribution determined by cascade impaction of exemplary formoterolfumarate co-suspension compositions prepared according to the presentdescription and stored for three months at 25° C./75% RH, withoutprotective overwrap, or at 40° C./75% RH with protective overwrap.

FIG. 22 is a graph, which depicts the chemical stability of exemplaryco-suspension compositions including formoterol fumarate as the activeagent. The results depicted in this figure allow comparison of thechemical stability of formoterol fumarate achieved in a co-suspensioncomposition formulated using crystalline formoterol fumarate with thechemical stability of suspension formulations prepared using spray driedformoterol fumarate.

FIG. 23 through FIG. 26 are electron micrographs of suspending particlesprepared from various different materials, with FIG. 23 providing amicrograph of trehalose suspending particles, FIG. 24 providing amicrograph of HP-β-cyclodextrin suspending particles, FIG. 25 providinga micrograph of Ficoll MP 70 suspending particles, and FIG. 26 providinga micrograph of inulin suspending particles.

FIG. 27 provides a graph that depicts the aerodynamic particle sizedistribution determined by cascade impaction of exemplary co-suspensioncompositions prepared according to the present description and includingglycopyrrolate active agent particles.

FIG. 28 provides a graph that depicts the aerodynamic particle sizedistribution determined by cascade impaction of exemplary co-suspensioncompositions prepared according to the present description and includingformoterol fumarate active agent particles.

FIG. 29 provides a graph that depicts the delivered dose uniformityachieved by ultra low-dose formoterol fumarate co-suspensioncompositions prepared according to the present description.

FIG. 30 provides graphs illustrating the particle size distribution ofglycopyrrolate (top) and formoterol (bottom) achieved by an exemplaryco-suspension compared to particle size distributions achieved byformulations including either glycopyrrolate or formoterol fumaratealone.

DETAILED DESCRIPTION

The present disclosure provides compositions, methods, and systems forrespiratory delivery of active agents via an MDI. In particularembodiments, the compositions, methods and systems described herein areadapted for respiratory delivery of active agents selected from along-acting muscarinic antagonist (“LAMA”) and a long-acting β₂adrenergic receptor agonist (“LABA”). In certain embodiments, the LAMAor LABA active agent may be potent or highly potent and, therefore,formulated at low concentrations and delivered in low doses. Thepharmaceutical compositions described herein may be formulated forpulmonary or nasal delivery via an MDI. The methods described hereininclude methods of stabilizing formulations including LAMA or LABAactive agents for respiratory delivery, as well as methods for pulmonarydelivery of LAMA and LABA active agents via an MDI. Also describedherein are methods for preparing an MDI for delivery of a LAMA or LABAactive agent.

In specific embodiments, the methods described herein include methodsfor treating a pulmonary disease or disorder amenable to treatment bydelivery of a LAMA or LABA active agent through an MDI. For example, andthe compositions, methods and systems described herein can be used totreat inflammatory or obstructive pulmonary diseases or conditions. Incertain embodiments, the compositions, methods and systems describedherein can be used to treat patients suffering from a disease ordisorder selected from asthma, chronic obstructive pulmonary disease(COPD), exacerbation of airways hyper reactivity consequent to otherdrug therapy, allergic rhinitis, sinusitis, pulmonary vasoconstriction,inflammation, allergies, impeded respiration, respiratory distresssyndrome, pulmonary hypertension, pulmonary vasoconstriction, and anyother respiratory disease, condition, trait, genotype or phenotype thatcan respond to the administration of a LAMA or LABA, alone or incombination with other therapies. In certain embodiments, thecompositions, systems and methods described herein can be used to treatpulmonary inflammation and obstruction associated with cystic fibrosis.As used herein, the terms “COPD” and “chronic obstructive pulmonarydisease” encompass chronic obstructive lung disease (COLD), chronicobstructive airway disease (COAD), chronic airflow limitation (CAL) andchronic obstructive respiratory disease (CORD) and include chronicbronchitis, bronchiectasis, and emphysema. As used herein, the term“asthma” refers to asthma of whatever type or genesis, including bothintrinsic (non-allergic) asthma and extrinsic (allergic) asthma, mildasthma, moderate asthma, severe asthma, bronchitic asthma,exercise-induced asthma, occupational asthma and asthma inducedfollowing bacterial infection. Asthma is also to be understood asembracing wheezy-infant syndrome.

It will be readily understood that the embodiments described herein areexemplary. The following more detailed description of variousembodiments is not intended to limit the scope of the presentdisclosure, but is merely representative of various embodiments.Moreover, the order of the steps or actions of the methods described inconnection with the embodiments disclosed herein may be changed by thoseskilled in the art without departing from the scope of the presentdisclosure. In other words, unless a specific order of steps or actionsis required for proper operation of the embodiment, the order or use ofspecific steps or actions may be modified.

I. DEFINITIONS

Unless specifically defined otherwise, the technical terms, as usedherein, have their normal meaning as understood in the art. Thefollowing terms are specifically defined for the sake of clarity.

The term “active agent” is used herein to include any agent, drug,compound, composition or other substance that may be used on, oradministered to a human or animal and is a LAMA or LABA. The term“active agent” may be used interchangeably with the terms, “drug,”“pharmaceutical,” “medicament,” “drug substance,” or “therapeutic.”

The terms “associate,” “associate with” or “association” refers to aninteraction or relationship between a chemical entity, composition, orstructure in a condition of proximity to a surface, such as the surfaceof another chemical entity, composition, or structure. The associationincludes, for example, adsorption, adhesion, covalent bonding, hydrogenbonding, ionic bonding and electrostatic attraction, Lifshitz-van derWaals interactions and polar interactions. The term “adhere” or“adhesion” is a form of association and is used as a generic term forall forces tending to cause a particle or mass to be attracted to asurface. “Adhere” also refers to bringing and keeping particles incontact with each other, such that there is substantially no visibleseparation between particles due to their different buoyancies in apropellant under normal conditions. In one embodiment, a particle thatattaches to or binds to a surface is encompassed by the term “adhere.”Normal conditions may include storage at room temperature or under anaccelerative force due to gravity. As described herein, active agentparticles may associate with suspending particles to form aco-suspension, where there is substantially no visible separationbetween the suspending particles and the active agent particles orflocculates thereof due to differences in buoyancy within a propellant.

“Suspending particles” refer to a material or combination of materialsthat is acceptable for respiratory delivery, and acts as a vehicle foractive agent particles. Suspending particles interact with the activeagent particles to facilitate repeatable dosing, delivery or transportof active agent to the target site of delivery, i.e., the respiratorytract. The suspending particles described herein are dispersed within asuspension medium including a propellant or propellant system, and canbe configured according to any shape, size or surface characteristicsuited to achieving a desired suspension stability or active agentdelivery performance. Exemplary suspending particles include particlesthat exhibit a particle size that facilitates respiratory delivery ofactive agent and have physical configurations suited to formulation anddelivery of the stabilized suspensions as described herein.

The term “co-suspension” refers to a suspension of two or more types ofparticles having different compositions within a suspension medium,wherein one type of particle associates at least partially with one ormore of the other particle types. The association leads to an observablechange in one or more characteristics of at least one of the individualparticle types suspended in the suspension medium. Characteristicsmodified by the association may include, for example, one or more of therate of aggregation or flocculation, the rate and nature of separation,i.e. sedimentation or creaming, density of a cream or sediment layer,adhesion to container walls, adhesion to valve components, and rate andthe level of dispersion upon agitation.

Exemplary methods for assessing whether a co-suspension is present caninclude the following: If one particle type has a pycnometric densitygreater than the propellant and another particle type has a pycnometricdensity lower than the propellant, a visual observation of the creamingor sedimentation behavior can be employed to determine the presence of aco-suspension. The term “pycnometric density” refers to the density of amaterial that makes up a particle, excluding voids within the particle.In one embodiment, the materials can be formulated or transferred into atransparent vial, typically a glass vial, for visual observation. Afterinitial agitation the vial is left undisturbed for a sufficient time forformation of a sediment or cream layer, typically 24 hours. If thesediment or cream layer is observed to be completely or mostly a uniformsingle layer, a co-suspension is present. The term “co-suspension”includes partial co-suspensions, where a majority of the at least twoparticle types associate with each other, however, some separation(i.e., less than a majority) of the at least two particle types may beobserved.

The exemplary co-suspension test may be performed at differentpropellant temperatures to accentuate the sedimentation or creamingbehavior of particle types with a density close to the propellantdensity at room temperature. If the different particle types have thesame nature of separation, i.e. all sediment or all cream, the presenceof a co-suspension can be determined by measuring other characteristicsof the suspension, such as rate of aggregation or flocculation, rate ofseparation, density of cream or sediment layer, adhesion to containerwalls, adhesion to valve components, and rate and level of dispersionupon agitation, and comparing them to the respective characteristics ofthe similarly suspended individual particle types. Various analyticalmethods generally known to those skilled in the art can be employed tomeasure these characteristics.

In the context of a composition containing or providing respirableaggregates, particles, drops, etc., such as compositions describedherein, the term “fine particle dose” or “FPD” refers to the dose,either in total mass or fraction of the nominal dose or metered dose,that is within a respirable range. The dose that is within therespirable range is measured in vitro to be the dose that depositsbeyond the throat stage of a cascade impactor, i.e., the sum of dosedelivered at stages 3 through filter in a Next Generation Impactoroperated at a flow rate of 30 l/min.

In the context of a composition containing or providing respirableaggregates, particles, drops, etc., such as compositions describedherein, the term “fine particle fraction” or “FPF” refers to theproportion of the delivered material relative to the delivered dose(i.e., the amount that exits the actuator of a delivery device, such asan MDI) that is within a respirable range. The amount of deliveredmaterial within the respirable range is measured in vitro as the amountof material that deposits beyond the throat stage of a cascade impactor,e.g., the sum of the material delivered at stages 3 through filter in aNext Generation Impactor operated at a flow rate of 30 l/min.

As used herein, the term “inhibit” refers to a measurable lessening ofthe tendency of a phenomenon, symptom or condition to occur or thedegree to which that phenomenon, symptom or condition occurs. The term“inhibit” or any form thereof, is used in its broadest sense andincludes minimize, prevent, reduce, repress, suppress, curb, constrain,restrict, slow progress of and the like.

“Mass median aerodynamic diameter” or “MMAD” as used herein refers tothe aerodynamic diameter of an aerosol below which 50% of the mass ofthe aerosol consists of particles with an aerodynamic diameter smallerthan the MMAD, with the MMAD being calculated according to monograph 601of the United States Pharmacopeia (“USP”).

When referred to herein, the term “optical diameter” indicates the sizeof a particle as measured by the Fraunhofer diffraction mode using alaser diffraction particle size analyzer equipped with a dry powderdispenser (e.g., Sympatec GmbH, Clausthal-Zellerfeld, Germany).

The term solution mediated transformation refers to the phenomenon inwhich a more soluble form of a solid material (i.e. particles with smallradius of curvature (a driving force for Ostwald ripening), or amorphousmaterial) dissolves and recrystallizes into the more stable crystal formthat can coexist in equilibrium with its saturated propellant solution.

A “patient” refers to an animal in which LAMA or LABA active agents willhave a therapeutic effect. In one embodiment, the patient is a humanbeing.

“Perforated microstructures” refer to suspending particles that includea structural matrix that exhibits, defines or comprises voids, pores,defects, hollows, spaces, interstitial spaces, apertures, perforationsor holes that allow the surrounding suspension medium to permeate, fillor pervade the microstructure, such as those materials and preparationsdescribed in U.S. Pat. No. 6,309,623 to Weers, et al. The primary formof the perforated microstructure is, generally, not essential, and anyoverall configuration that provides the desired formulationcharacteristics is contemplated herein. Accordingly, in one embodiment,the perforated microstructures may comprise approximately sphericalshapes, such as hollow, suspending, spray-dried microspheres. However,collapsed, corrugated, deformed or fractured particulates of any primaryform or aspect ratio may also be compatible.

As is true of suspending particles described herein, perforatedmicrostructures may be formed of any biocompatible material that doesnot substantially degrade or dissolve in the selected suspension medium.While a wide variety of materials may be used to form the particles, insome embodiments, the structural matrix is associated with, or includes,a surfactant such as, a phospholipid or fluorinated surfactant. Althoughnot required, the incorporation of a compatible surfactant in theperforated microstructure or, more generally, the suspending particles,can improve the stability of the respiratory dispersions, increasepulmonary deposition and facilitate the preparation of the suspension.

The term “suspension medium” as uses herein refers to a substanceproviding a continuous phase within which active agent particles andsuspending particles can be dispersed to provide a co-suspensionformulation. The suspension medium used in co-suspension formulationsdescribed herein includes propellant. As used herein, the term“propellant” refers to one or more pharmacologically inert substanceswhich exert a sufficiently high vapor pressure at normal roomtemperature to propel a medicament from the canister of an MDI to apatient on actuation of the MDI's metering valve. Therefore, the term“propellant” refers to both a single propellant and to a combination oftwo or more different propellants forming a “propellant system.”

The term “respirable” generally refers to particles, aggregates, drops,etc. sized such that they can be inhaled and reach the airways of thelung.

When used to refer to co-suspension compositions described herein, theterms “physical stability” and “physically stable” refer to acomposition that is resistant to one or more of aggregation,flocculation, and particle size changes due to solution mediatedtransformations and is capable of substantially maintaining the MMAD ofsuspending particles and the fine particle dose. In one embodiment,physical stability may be evaluated through subjecting compositions toaccelerated degradation conditions, such as by temperature cycling asdescribed herein.

When referring to active agents, the term “potent” indicates activeagents that are therapeutically effective at or below doses ranging fromabout 0.01 mg/kg to about 1 mg/kg. Typical doses of potent active agentsgenerally range from about 100 μg to about 100 mg.

When referring to active agents, the term “highly potent” indicatesactive agents that are therapeutically effective at or below doses ofabout 10 μg/kg. Typical doses of highly potent active agents generallyrange up to about 100 μg.

The terms “suspension stability” and “stable suspension” refer tosuspension formulations capable of maintaining the properties of aco-suspension of active agent particles and suspending particles over aperiod of time. In one embodiment, suspension stability may be measuredthrough delivered dose uniformity achieved by co-suspension compositionsdescribed herein.

The term “substantially insoluble” means that a composition is eithertotally insoluble in a particular solvent or it is poorly soluble inthat particular solvent. The term “substantially insoluble” means that aparticular solute has a solubility of less than one part per 100 partssolvent. The term “substantially insoluble” includes the definitions of“slightly soluble” (from 100 to 1000 parts solvent per 1 part solute),“very slightly soluble” (from 1000 to 10,000 parts solvent per 1 partsolute) and “practically insoluble” (more than 10,000 parts solvent per1 part solute) as given in Table 16-1 of Remington: The Science andPractice of Pharmacy, 21st ed. Lippincott, Williams & Wilkins, 2006, p.212.

The term “surfactant,” as used herein, refers to any agent whichpreferentially adsorbs to an interface between two immiscible phases,such as the interface between water and an organic polymer solution, awater/air interface or organic solvent/air interface. Surfactantsgenerally possess a hydrophilic moiety and a lipophilic moiety, suchthat, upon adsorbing to microparticles, they tend to present moieties tothe continuous phase that do not attract similarly-coated particles,thus reducing particle agglomeration. In some embodiments, surfactantsmay also promote adsorption of a drug and increase bioavailability ofthe drug.

A “therapeutically effective amount” is the amount of compound whichachieves a therapeutic effect by inhibiting a disease or disorder in apatient or by prophylactically inhibiting or preventing the onset of adisease or disorder. A therapeutically effective amount may be an amountwhich relieves to some extent one or more symptoms of a disease ordisorder in a patient; returns to normal either partially or completelyone or more physiological or biochemical parameters associated with orcausative of the disease or disorder; and/or reduces the likelihood ofthe onset of the disease of disorder.

The terms “chemically stable” and “chemical stability” refer toco-suspension formulations wherein the individual degradation productsof active agent remain below the limits specified by regulatoryrequirements during the shelf life of the product for human use (e.g.,1% of total chromatographic peak area per ICH guidance Q3B(R2)) andthere is acceptable mass balance (e.g., as defined in ICH guidance Q1E)between active agent assay and total degradation products.

II. PHARMACEUTICAL COMPOSITIONS

The compositions described herein are co-suspensions that include asuspension medium including a propellant, LAMA or LABA active agentparticles, and suspending particles. Of course, if desired, thecompositions described herein may include one or more additionalconstituents. Moreover, variations and combinations of components of thecompositions described herein may be used. For example, two or morespecies of suspending particles may be used in compositions for theformulation and delivery of a selected LAMA or LABA active agent.Alternatively, for example, the compositions described herein mayinclude two or more species of active agent particles. In certain suchembodiments, the compositions may include LAMA or LABA active agentparticles co-suspended with suspending particles, wherein, in additionto the active agent material included in the active agent particles, atleast some of the suspending particles incorporate the selected LAMA orLABA active agent. Even further, if desired, the compositions describedherein may include two or more different species of particles containingthe selected LAMA or LABA active agent in combination with two or moredifferent species of suspending particles.

It has been found that, in formulations according to the presentdescription, active agent particles exhibit an association with thesuspending particles such that separation of the active agent particlesfrom the suspending particles is substantially prevented, resulting inco-location of active agent particles and suspending particles withinthe suspension medium. Generally, due to density differences betweendistinct species of particles and the medium within which they aresuspended (e.g., a propellant or propellant system), buoyancy forcescause creaming of particles with lower density than the propellant andsedimentation of particles with higher density than the propellant.Therefore, in suspensions that include a mixture of particles that varyin their densities, the sedimentation or creaming behavior of each typeof particle may vary and may lead to separation of the differentparticle types within the propellant.

However, the combinations of propellant, active agent particles andsuspending particles described herein provide co-suspensions wherein theactive agent particles and suspending particles co-locate within thepropellant (i.e., the active agent particles associate with thesuspending particles such that suspending particles and active agentparticles do not exhibit substantial separation relative to each other,such as by differential sedimentation or creaming, even after a timesufficient for the formation of a cream or sediment layer). Inparticular embodiments, for example, the compositions described hereinform co-suspensions wherein the suspending particles remain associatedwith active agent particles when subjected to buoyancy forces amplifiedby temperature fluctuations and/or centrifugation at accelerations up toan over, for example, 1 g, 10 g, 35 g, 50 g, and 100 g. However, theco-suspensions described herein need not be defined by or limited to aspecific threshold force of association. For example, a co-suspension ascontemplated herein may be successfully achieved where the active agentparticles associate with the suspending particles such that there is nosubstantial separation of active agent particles and suspendingparticles within the continuous phase formed by the suspension mediumunder typical patient use conditions.

Co-suspension compositions according to the present description providedesirable formulation and delivery characteristics for LAMA and LABAactive agents. For example, in certain embodiments, when present withinan MDI canister, co-suspensions as described herein can inhibit orreduce one or more of the following: flocculation of active agentmaterial; differential sedimentation or creaming of active agentparticles and suspending particles; solution mediated transformation ofactive agent material; and loss of active agent to the surfaces of thecontainer closure system, in particular the metering valve components.In addition, compositions as described herein provide chemical stabilityfor the active agents contained therein. Such qualities work to achieveand preserve aerosol performance as the co-suspension is delivered froman MDI such that desirable fine particle fraction, fine particle doseand delivered dose uniformity characteristics are achieved andsubstantially maintained throughout emptying of an MDI canister withinwhich the co-suspension composition is contained. Additionally, asillustrated by embodiments detailed herein, co-suspensions according tothe present description can provide a stable formulation that providesconsistent dosing and respiratory delivery characteristics for LAMA andLABA active agents, while utilizing a relatively simple HFA suspensionmedium that does not require modification by the addition of, forexample, cosolvents, antisolvents, solubilizing agents or adjuvants.

Providing a co-suspension according to the present description may alsosimplify formulation, delivery and dosing of LAMA and LABA activeagents. Without being bound by a particular theory, it is thought thatby achieving a co-suspension of active agent particles and suspendingparticles, the delivery and dosing of active agent contained within sucha dispersion may be substantially controlled through control of thesize, composition, morphology and relative amount of the suspendingparticles, and less dependent upon the size and morphology of the activeagent particles.

Accordingly, the pharmaceutical compositions disclosed herein providefor delivery of LAMA and LABA active agents from an MDI. Delivery of theco-suspension compositions described herein provides desirablepharmacokinetic and pharmacodynamic characteristics, and MDI delivery ofthe pharmaceutical compositions described herein is suitable fortreating patients suffering from an inflammatory or obstructivepulmonary disease or condition that responds to the administration of aLAMA or LABA active agent. In particular embodiments, the pharmaceuticalcompositions described herein may be used in treating a disease orcondition selected from asthma, COPD, exacerbation of airways hyperreactivity consequent to other drug therapy, allergic rhinitis,sinusitis, pulmonary vasoconstriction, inflammation, allergies, impededrespiration, respiratory distress syndrome, pulmonary hypertension,pulmonary vasoconstriction, emphysema, and any other respiratorydisease, condition, trait, genotype or phenotype that can respond to theadministration of a LAMA or LABA, alone or in combination with othertherapies. In certain embodiments, the compositions, systems and methodsdescribed herein can be used to treat pulmonary inflammation andobstruction associated with cystic fibrosis.

(i) Suspension Medium

The suspension medium included in a composition described hereinincludes one or more propellants. In general, suitable propellants foruse as suspension mediums are those propellant gases that can beliquefied under pressure at room temperature, and upon inhalation ortopical use, are safe and toxicologically innocuous. Additionally, it isdesirable that the selected propellant be relatively non-reactive withthe suspending particles or active agent particles. Exemplary compatiblepropellants include hydrofluoroalkanes (HFAs), perfluorinated compounds(PFCs), and chlorofluorocarbons (CFCs).

Specific examples of propellants that may be used to form the suspensionmedium of the co-suspensions disclosed herein include1,1,1,2-tetrafluoroethane (CF₃CH₂F) (HFA-134a),1,1,1,2,3,3,3-heptafluoro-n-propane (CF₃CHFCF₃) (HFA-227),perfluoroethane, monochloro-fluoromethane, 1,1 difluoroethane, andcombinations thereof. Even further, suitable propellants include, forexample: short chain hydrocarbons; C₁₋₄ hydrogen-containingchlorofluorocarbons such as CH₂ClF, CCl₂FCHClF, CF₃CHClF, CHF₂CClF₂,CHClFCHF₂, CF₃CH₂Cl, and CClF₂CH₃; C₁₋₄ hydrogen-containingfluorocarbons (e.g., HFAs) such as CHF₂CHF₂, CF₃CH₂F, CHF₂CH₃, andCF₃CHFCF₃; and perfluorocarbons such as CF₃CF₃ and CF₃CF₂CF₃.

Specific fluorocarbons, or classes of fluorinated compounds, that may beused as suspension media include, but are not limited to, fluoroheptane,fluorocycloheptane, fluoromethylcycloheptane, fluorohexane,fluorocyclohexane, fluoropentane, fluorocyclopentane,fluoromethylcyclopentane, fluorodimethyl-cyclopentanes,fluoromethylcyclobutane, fluorodimethylcyclobutane,fluorotrimethyl-cyclobutane, fluorobutane, fluorocyclobutane,fluoropropane, fluoroethers, fluoropolyethers and fluorotriethylamines.These compounds may be used alone or in combination with more volatilepropellants.

In addition to the aforementioned fluorocarbons and hydrofluoroalkanes,various exemplary chlorofluorocarbons and substituted fluorinatedcompounds may also be used as suspension media. In this respect, FC-11(CCl₃F), FC-11B1 (CBrCl₂F), FC-11B2 (CBr₂ClF), FC12B2 (CF₂Br₂), FC21(CHCl₂F), FC21B1 (CHBrClF), FC-21B2 (CHBr₂F), FC-31B1 (CH₂BrF), FC113A(CCl₃CF₃), FC-122 (CClF₂CHCl₂), FC-123 (CF₃CHCl₂), FC-132 (CHClFCHClF),FC-133 (CHClFCHF₂), FC-141 (CH₂ClCHClF), FC-141B (CCl₂FCH₃), FC-142(CHF₂CH₂Cl), FC-151 (CH₂FCH₂Cl), FC-152 (CH₂FCH₂F), FC-1112 (CClF═CClF),FC-1121 (CHCl═CFCl) and FC-1131 (CHCl═CHF) may also be used, whilerecognizing the possible attendant environmental concerns. As such, eachof these compounds may be used, alone or in combination with othercompounds (i.e., less volatile fluorocarbons) to form the stabilizedsuspensions disclosed herein.

In some embodiments, the suspension medium may be formed of a singlepropellant. In other embodiments, a combination of propellants (a“propellant system”) may be used to form the suspension medium. In someembodiments, relatively volatile compounds may be mixed with lower vaporpressure components to provide suspension media having specifiedphysical characteristics selected to improve stability or enhance thebioavailability of the dispersed active agent. In some embodiments, thelower vapor pressure compounds will comprise fluorinated compounds (e.g.fluorocarbons) having a boiling point greater than about 25° C. In someembodiments, lower vapor pressure fluorinated compounds for use in thesuspension medium may include perfluorooctylbromide C₈F₁₇Br (PFOB orperflubron), dichlorofluorooctane C₅F₁₆Cl₂, perfluorooctylethaneC₈F₁₇C₂H₅ (PFOE), perfluorodecylbromide C₁₀F₂₁Br (PFDB) orperfluorobutylethane C₄F₉C₂H₅. In certain embodiments, these lower vaporpressure compounds are present in a relatively low level. Such compoundsmay be added directly to the suspension medium or may be associated withthe suspending particles.

The suspension medium included in compositions as described herein maybe formed of a propellant or propellant system that is substantiallyfree of additional materials, including, for example, antisolvents,solubilizing agents, cosolvents or adjuvants. For example, in someembodiments, the suspension medium may be formed of a non-CFC propellantor propellant system, such as an HFA propellant or propellant system,that is substantially free of additional materials. Such embodimentssimplify the formulation and manufacture of pharmaceutical compositionssuited for respiratory delivery of a LAMA or LABA active agent.

However, in other embodiments, depending on the selection of propellant,the properties of the suspending particles, or the nature of activeagent to be delivered, the suspension medium utilized may includematerials in addition to the propellant or propellant system. Suchadditional materials may include, for example, one or more of anappropriate antisolvent, solubilizing agent, cosolvent or adjuvant toadjust, for example, the vapor pressure of the formulation or thestability, or solubility of suspended particles. For example, propane,ethanol, isopropyl alcohol, butane, isobutane, pentane, isopentane or adialkyl ether, such as dimethyl ether, may be incorporated with thepropellant in the suspension medium. Similarly, the suspension mediummay contain a volatile fluorocarbon. In other embodiments, one or bothof polyvinylpyrrolidone (“PVP”) or polyethylene glycol (“PEG”) may beadded to the suspension medium. Adding PVP or PEG to the suspensionmedium may achieve one or more desired functional characteristics, andin one example, PVP or PEG may be added to the suspension medium as acrystal growth inhibitor. In general, where a volatile cosolvent oradjuvant is used, such an adjuvant or cosolvent may be selected fromknown hydrocarbon or fluorocarbon materials and may account for up toabout 1% w/w of the suspension medium. For example, where a cosolvent oradjuvant is incorporated in the suspension medium, the cosolvent oradjuvant may comprise less than about 0.01%, 0.1%, or 0.5% w/w of thesuspension medium. Where PVP or PEG are included in the suspensionmedium, such constituents may be included at up to about 1% w/w, or theymay comprise less than about 0.01%, 0.1%, or 0.5% w/w of the suspensionmedium.

(ii) Active Agent Particles

The active agent particles included in the co-suspensions describedherein are formed to be capable of being dispersed and suspended withinthe suspension medium and are sized to facilitate delivery of respirableparticles from the co-suspension. In one embodiment, therefore, theactive agent particles are provided as a micronized material wherein atleast 90% of the active agent particle material by volume exhibits anoptical diameter of about 7 μm or less. In other embodiments, the activeagent particles are provided as a micronized material wherein at least90% of the active agent particles by volume exhibit an optical diameterselected from a range of about 6 μm to about 1 μm, about 5 μm to about 2μm, and about 4 μm to about 3 μm. In further embodiments, the activeagent particles are provided as a micronized material wherein at least90% of the active agent particles by volume exhibit an optical diameterselected from 6 μm or less, 5 μm or less, and 4 μm or less. In anotherembodiment, the active agent particles are provided as a micronizedmaterial wherein at least 50% of the active agent particle material byvolume exhibits an optical diameter of about 5 μm or less. In otherembodiments, the active agent particles are provided as a micronizedmaterial wherein at least 50% of the active agent particles by volumeexhibit an optical diameter selected from a range of about 4 μm to about1 μm, about 3 μm to about 1 μm, and about 2.5 μm to about 1 μm. Inanother embodiment, the active agent particles are provided as amicronized material wherein at least 50% of the active agent particlesby volume exhibit an optical diameter selected from 4 μm or less, 3 μmor less, and 2 μm or less.

In specific embodiments, the active agent material used as or to formthe active agent particles may be entirely or substantially crystalline,i.e., a majority of the active agent molecules are arranged in aregularly repeating pattern, over a long range or external face planes.In another embodiment, the active agent particles may be present in bothcrystal and amorphous states. In yet another embodiment, the activeagent particles may be present in substantially an amorphous state,i.e., the active agent particles are overall noncrystalline in natureand do not have a regularly repeating arrangement of moleculesmaintained over along range. Suitable excipients for formulation ofactive agent particles include those described herein in associationwith the suspending particles. In specific embodiments, for example,active agent particles may be formulated with one or more of the lipid,phospholipid, carbohydrate, amino acid, organic salt, peptide, protein,alditols, synthetic or natural polymer, or surfactant materials asdescribed, for example, in association with the suspending particles. Inother embodiments, the active agent particles are formed solely frommicronized active agent material.

Because the compositions disclosed enable the formulation andreproducible delivery of very low doses of active agents, in certainembodiments, the active agents included in the compositions describedherein may be selected from one or more potent or highly potent activeagents. For example, in certain embodiments, the compositions describedherein may include a potent active agent that is delivered at a singleadministration dose selected from between about 100 μg and about 100 mgper dose, about 100 μg and about 10 mg per dose, and about 100 μg and 1mg per dose. In other embodiments, the compositions described herein mayinclude a potent or highly potent active agent that is delivered at adose selected from up to about 80 μg per single administration dose, upto about 40 μg per single administration dose, up to about 20 μg persingle administration dose, up to about 10 μg per single administrationdose or between about 10 μg and about 100 μg per single administrationdose. Additionally, in certain embodiments, the compositions describedherein may include a highly potent active agent delivered at a doseselected from between about 0.1 and about 2 μg per single administrationdose, about 0.1 and about 1 μg per single administration dose, and about0.1 and about 0.5 μg per single administration dose.

In certain embodiments, the active agent included in the compositionsdescribed herein is a LAMA active agent. Where the compositions includea LAMA active agent, in particular embodiments, the LAMA active agentmay be selected from, for example, glycopyrrolate, dexipirronium,tiotropium, trospium, aclidinium, darotropium, including anypharmaceutically acceptable salts, esters, isomers or solvates thereof.

Glycopyrrolate can be used to treat inflammatory or obstructivepulmonary diseases and disorders such as, for example, those describedherein. As an anticholinergic, glycopyrrolate acts as a bronchodilatorand provides an antisecretory effect, which is a benefit for use in thetherapy of pulmonary diseases and disorders characterized by increasedmucus secretions. Glycopyrrolate is a quaternary ammonium salt. Whereappropriate, glycopyrrolate may be used in the form of salts (e.g.alkali metal or amine salts, or as acid addition salts) or as esters oras solvates (hydrates). Additionally, the glycopyrrolate may be in anycrystalline form or isomeric form or mixture of isomeric forms, forexample a pure enantiomer, a mixture of enantiomers, a racemate or amixture thereof. In this regard, the form of glycopyrrolate may beselected to optimize the activity and/or stability of glycopyrrolateand/or to minimize the solubility of glycopyrrolate in the suspensionmedium. Suitable counter ions are pharmaceutically acceptable counterions including, for example, fluoride, chloride, bromide, iodide,nitrate, sulfate, phosphate, formate, acetate, trifluoroacetate,propionate, butyrate, lactate, citrate, tartrate, malate, maleate,succinate, benzoate, p-chlorobenzoate, diphenyl-acetate ortriphenylacetate, o-hydroxybenzoate, p-hydroxybenzoate,1-hydroxynaphthalene-2-carboxylate, 3-hydroxynaphthalene-2-carboxylate,methanesulfonate and benzenesulfonate. In particular embodiments of thecompositions described herein, the bromide salt of glycopyrrolate,namely3-[(cyclopentyl-hydroxyphenylacetyl)oxy]-1,1-dimethylpyrrolidiniumbromide, is used and can be prepared according to the procedures set outin U.S. Pat. No. 2,956,062.

Where the compositions described herein include glycopyrrolate, incertain embodiments, the compositions may include sufficientglycopyrrolate to provide a target delivered dose selected from betweenabout 10 μg and about 200 μg per actuation of an MDI, about 15 μg andabout 150 μg per actuation of an MDI, and about 18 μg and 144 μg peractuation of an MDI. In other such embodiments, the formulations includesufficient glycopyrrolate to provide a dose selected from up to about200 μg, up to about 150 μg, up to about 75 μg, up to about 40 μg, or upto about 20 μg per actuation. In yet further embodiments, theformulations include sufficient glycopyrrolate to provide a doseselected from about 18 μg per actuation, 36 μg per actuation, or about72 μg per actuation. In order to achieve targeted delivered doses asdescribed herein, where compositions described herein includeglycopyrrolate as the active agent, in specific embodiments, the amountof glycopyrrolate included in the compositions may be selected from, forexample, between about 0.04 mg/ml and about 2.25 mg/ml.

In other embodiments, tiotropium, including any pharmaceuticallyacceptable salts, esters, isomers or solvates thereof, may be selectedas a LAMA active agent for inclusion in a composition as describedherein. Tiotropium is a known, long-acting anticholinergic drug suitablefor use in treating diseases or disorders associated with pulmonaryinflammation or obstruction, such as those described herein. Tiotropium,including crystal and pharmaceutically acceptable salt forms oftiotropium, is described, for example, in U.S. Pat. No. 5,610,163, U.S.Pat. No. RE39820, U.S. Pat. No. 6,777,423, and U.S. Pat. No. 6,908,928.Where the compositions described herein include tiotropium, in certainembodiments, the compositions may include sufficient tiotropium toprovide a delivered dose selected from between about 2.5 μg and about 50μg, about 4 μg and about 25 μg, about 2.5 μg and about 20 μg, about 10μg and about 20 μg, and about 2.5 μg and about 10 μg per actuation of anMDI. In other such embodiments, the formulations include sufficienttiotropium to provide a delivered dose selected from up to about 50 μg,up to about 20 μg, up to about 10 μg, up to about 5 μg, or up to about2.5 μg per actuation of an MDI. In yet further embodiments, theformulations include sufficient tiotropium to provide a delivered doseselected from about 3 μg, 6 μg, 9 μg, 18 μg, and 36 μg per actuation ofthe MDI. In order to achieve delivered doses as described herein, wherecompositions described herein include tiotropium as the active agent, inspecific embodiments, the amount of tiotropium included in thecompositions may be selected from, for example, between about 0.01 mg/mland about 0.5 mg/ml.

In certain embodiments, the compositions described herein include a LABAactive agent. In such embodiments, a LABA active agent can be selectedfrom, for example, bambuterol, clenbuterol, formoterol, salmeterol,carmoterol, milveterol, indacaterol, and saligenin- or indole-containingand adamantyl-derived β₂ agonists, and any pharmaceutically acceptablesalts, esters, isomers or solvates thereof. In certain such embodiments,formoterol is selected as the LABA active agent. Formoterol can be usedto treat inflammatory or obstructive pulmonary diseases and disorderssuch as, for example, those described herein. Formoterol has thechemical name(±)-2-hydroxy-5-[(1RS)-1-hydroxy-2-[[(1RS)-2-(4-methoxyphenyl)-1-methylethyl]-amino]ethyl]formanilide, and is commonly used in pharmaceutical compositions as theracemic fumarate dihydrate salt. Where appropriate, formoterol may beused in the form of salts (e.g. alkali metal or amine salts or as acidaddition salts) or as esters or as solvates (hydrates). Additionally,the formoterol may be in any crystalline form or isomeric form ormixture of isomeric forms, for example a pure enantiomer, a mixture ofenantiomers, a racemate or a mixture thereof. In this regard, the formof formoterol may be selected to optimize the activity and/or stabilityof formoterol and/or to minimize the solubility of formoterol in thesuspension medium. Pharmaceutically acceptable salts of formoterolinclude, for example, salts of inorganic acids such as hydrochloric,hydrobromic, sulfuric and phosphoric acids, and organic acids such asfumaric, maleic, acetic, lactic, citric, tartaric, ascorbic, succinic,glutaric, gluconic, tricarballylic, oleic, benzoic, p-methoxybenzoic,salicylic, o- and p-hydroxybenzoic, p-chlorobenzoic, methanesulfonic,p-toluenesulfonic and 3-hydroxy-2-naphthalene carboxylic acids. Hydratesof formoterol are described, for example, in U.S. Pat. No. 3,994,974 andU.S. Pat. No. 5,684,199. Specific crystalline forms of formoterol andother β₂ adrenergic receptor agonists are described, for example, inWO95/05805, and specific isomers of formoterol are described in U.S.Pat. No. 6,040,344.

In specific embodiments, the formoterol material utilized to form theformoterol particles is formoterol fumarate, and in one such embodiment,the formoterol fumarate is present in the dihydrate form. Where thecompositions described herein include formoterol, in certainembodiments, the compositions described herein may include formoterol ata concentration that achieves a targeted delivered dose selected frombetween about 1 μg and about 30 μg, about 1 μg and about 10 μg, about 2μg and 5 μg, about 2 μg and about 10 μg, about 5 μg and about 10 μg, and3 μg and about 30 μg per actuation of an MDI. In other embodiments, thecompositions described herein may include formoterol in an amountsufficient to provide a targeted delivered dose selected from up toabout 30 μg, up to about 10 μg, up to about 5 μg, up to about 2.5 μg, upto about 2 μg, or up to about 1.5 μg per actuation. In order to achievetargeted delivered doses as described herein, where compositionsdescribed herein include formoterol as the active agent, in specificembodiments, the amount of formoterol included in the compositions maybe selected from, for example, between about 0.01 mg/ml and about 1mg/ml, between about 0.01 mg/ml and about 0.5 mg/ml, and between about0.03 mg/ml and about 0.4 mg/ml.

Where the pharmaceutical co-suspension compositions described hereininclude a LABA active agent, in certain embodiments, the active agentmay be salmeterol, including any pharmaceutically acceptable salts,esters, isomers or solvates thereof. Salmeterol can be used to treatinflammatory or obstructive pulmonary diseases and disorders such as,for example, those described herein. Salmeterol, pharmaceuticallyacceptable salts of salmeterol, and methods for producing the same aredescribed, for example, in U.S. Pat. No. 4,992,474, U.S. Pat. No.5,126,375, and U.S. Pat. No. 5,225,445.

Where salmeterol is included as a LABA active agent, in certainembodiments, the compositions described herein may include salmeterol ata concentration that achieves a delivered dose selected from betweenabout 2 μg and about 120 μg, about 4 μg and about 40 μg, about 8 μg and20 μg, about 8 μg and about 40 μg, about 20 μg and about 40 μg, and 12μg and about 120 μg per actuation of an MDI. In other embodiments, thecompositions described herein may include salmeterol in an amountsufficient to provide a delivered dose selected from up to about 120 μg,up to about 40 μg, up to about 20 μg, up to about 10 μg, up to about 8μg, or up to about 6 μg per actuation of an MDI. In order to achievetargeted delivered doses as described herein, where compositionsdescribed herein include salmeterol as the active agent, in specificembodiments, the amount of salmeterol included in the compositions maybe selected from, for example, between about 0.04 mg/ml and about 4mg/ml, between about 0.04 mg/ml and about 2.0 mg/ml, and between about0.12 mg/ml and about 0.8 mg/ml. For example, the compositions describedherein may include sufficient salmeterol to provide a target delivereddose selected from between about 4 μg and about 120 μg, about 20 μg andabout 100 μg, and between about 40 μg and about 120 μg per actuation ofan MDI. In still other embodiments, the compositions described hereinmay include sufficient salmeterol to provide a targeted delivered doseselected from up to about 100 μg, up to about 40 μg, or up to about 15μg per actuation of an MDI.

Though the active agent material included in the compositions describedherein may be amorphous or substantially amorphous, in specificembodiments, the active agent material used as or in the formation ofthe active agent particles included in the compositions described hereinis substantially or entirely crystalline. Active agent material that issubstantially or entirely crystalline may be selected to improve thechemical stability of the LAMA or LABA active agent when formulated inthe compositions described herein. Therefore, in specific embodiments,the active agent material included in the compositions described hereinis a micronized, crystalline LAMA material. In one such embodiment, theactive agent particles are formed solely of micronized, crystalline LAMAmaterial, such as a micronized crystalline material selected fromglycopyrrolate, dexipirronium, tiotropium, trospium, aclidinium,darotropium, and any pharmaceutically acceptable salts, esters orsolvates thereof. In other specific embodiments, the active agentmaterial included in the compositions described herein is a micronized,crystalline LABA material. In one such embodiment, the active agentparticles are formed solely of micronized, crystalline LABA material,such as a micronized crystalline material selected from bambuterol,clenbuterol, formoterol, salmeterol, carmoterol, milveterol,indacaterol, and saligenin- or indole-containing and adamantyl-derivedβ₂ agonists, and any pharmaceutically acceptable salts, esters orsolvates thereof.

Any suitable process may be employed to achieve micronized active agentmaterial as or in the formulation of the active agent particles includedin the compositions described herein. A variety of processes may be usedto create active agent particles suitable for use in the co-suspensionformulations described herein, including, but not limited tomicronization by milling or grinding processes, crystallization orrecrystallization processes, and processes using precipitation fromsupercritical or near-supercritical solvents, spray drying, spray freezedrying, or lyophilization. Patent references teaching suitable methodsfor obtaining micronized active agent particles are described, forexample, in U.S. Pat. No. 6,063,138, U.S. Pat. No. 5,858,410, U.S. Pat.No. 5,851,453, U.S. Pat. No. 5,833,891, U.S. Pat. No. 5,707,634, andInternational Patent Publication No. WO 2007/009164. Where the activeagent particles include active agent material formulated with one ormore excipient or adjuvant, micronized active agent particles can beformed using one or more of the preceding processes and such processescan be utilized to achieve active agent particles having a desired sizedistribution and particle configuration.

(iii) Suspending Particles

The suspending particles included in the co-suspension compositionsdescribed herein work to facilitate stabilization and delivery of theactive agent included in the compositions. Though various forms ofsuspending particles may be used, the suspending particles are typicallyformed from pharmacologically inert material that is acceptable forinhalation and is substantially insoluble in the propellant selected.Generally, the majority of suspending particles are sized within arespirable range. In particular embodiments, therefore, the MMAD of thesuspending particles will not exceed about 10 μm but is not lower thanabout 500 nm. In an alternative embodiment, the MMAD of the suspendingparticles is between about 5 μm and about 750 nm. In yet anotherembodiment, the MMAD of the suspending particles is between about 1 μmand about 3 μm. When used in an embodiment for nasal delivery from anMDI, the MMAD of the suspending particles is between 10 μm and 50 μm.

In order to achieve respirable suspending particles within the MMADranges described, the suspending particles will typically exhibit avolume median optical diameter between about 0.2 μm and about 50 μm. Inone embodiment, the suspending particles exhibit a volume median opticaldiameter that does not exceed about 25 μm. In another embodiment, thesuspending particles exhibit a volume median optical diameter selectedfrom between about 0.5 μm and about 15 μm, between about 1.5 μm andabout 10 μm, and between about 2 μm and about 5 μm.

The concentration of suspending particles included in a compositionaccording to the present description can be adjusted, depending on, forexample, the amount of active agent particles and suspension mediumused. In one embodiment, the suspending particles are included in thesuspension medium at a concentration selected from about 1 mg/ml toabout 15 mg/ml, about 3 mg/ml to about 10 mg/ml, 5 mg/ml to about 8mg/ml, and about 6 mg/ml. In another embodiment, the suspendingparticles are included in the suspension medium at a concentration of upto about 30 mg/ml. In yet another embodiment, the suspending particlesare included in the suspension medium at a concentration of up to about25 mg/ml.

The relative amount of suspending particles to active agent particles isselected to achieve a co-suspension as contemplated herein. Aco-suspension composition may be achieved where the amount of suspendingparticles, as measured by mass, exceeds that of the active agentparticles. For example, in specific embodiments, the ratio of the totalmass of the suspending particles to the total mass of active agentparticles may be between about 3:1 and about 15:1, or alternatively fromabout 2:1 and 8:1. Alternatively, the ratio of the total mass of thesuspending particles to the total mass of active agent particles may beabove about 1, such as up to about 1.5, up to about 5, up to about 10,up to about 15, up to about 17, up to about 20, up to about 30, up toabout 40, up to about 50, up to about 60, up to about 75, up to about100, up to about 150, and up to about 200, depending on the nature ofthe suspending particles and active agent particles used. In furtherembodiments, the ratio of the total mass of the suspending particles tothe total mass of the active agent particles may be selected frombetween about 10 and about 200, between about 60 and about 200, betweenabout 15 and about 60, between about 15 and about 170, between about 15and about 60, about 16, about 60, and about 170.

In other embodiments, the amount of suspending particles, as measured bymass, is less than that of the active agent particles. For example, inparticular embodiments, the mass of the suspending particles may be aslow as 20% of the total mass of the active agent particles. However, insome embodiments, the total mass of the suspending particles may alsoapproximate or equal the total mass of the active agent particles.

Suspending particles suitable for use in the compositions describedherein may be formed of one or more pharmaceutically acceptablematerials or excipients that are suitable for inhaled delivery and donot substantially degrade or dissolve in the suspension medium. In oneembodiment, perforated microstructures, as defined herein, may be usedas the suspending particles. Exemplary excipients that may be used inthe formulation of suspending particles described herein include but arenot limited to (a) carbohydrates, e.g., monosaccharides such asfructose, galactose, glucose, D-mannose, sorbose, and the like;disaccharides, such as sucrose, lactose, trehalose, cellobiose, and thelike; cyclodextrins, such as 2-hydroxypropyl-β-cyclodextrin; andpolysaccharides, such as raffinose, maltodextrins, dextrans, starches,chitin, chitosan, inulin, and the like; (b) amino acids, such asalanine, glycine, arginine, aspartic acid, glutamic acid, cysteine,lysine, leucine, isoleucine, valine, and the like; (c) metal and organicsalts prepared from organic acids and bases, such as sodium citrate,sodium ascorbate, magnesium gluconate, sodium gluconate, tromethaminhydrochloride, and the like; (d) peptides and proteins such asaspartame, trileucine, human serum albumin, collagen, gelatin, and thelike; (e) alditols, such as mannitol, xylitol, and the like; (f)synthetic or natural polymers or combinations thereof, such aspolylactides, polylactide-glycolides, cyclodextrins, polyacrylates,methylcellulose, carboxymethylcellulose, polyvinyl alcohols,polyanhydrides, polylactams, polyvinyl pyrrolidones, hyaluronic acid,polyethylene glycols; and (g) surfactants including fluorinated andnonfluorinated compounds such as saturated and unsaturated lipids,nonionic detergents, nonionic block copolymers, ionic surfactants andcombinations thereof. In particular embodiments, suspending particlesmay include a calcium salt, such as calcium chloride, as described, forexample, in U.S. Pat. No. 7,442,388.

Additionally, phospholipids from both natural and synthetic sources maybe used in preparing suspending particles suitable for use in thecompositions described herein. In particular embodiments, thephospholipid chosen will have a gel to liquid crystal phase transitionof greater than about 40° C. Exemplary phospholipids are relatively longchain (i.e., C₁₆-C₂₂) saturated lipids and may comprise saturatedphospholipids, such as saturated phosphatidylcholines having acyl chainlengths of 16 C or 18 C (palmitoyl and stearoyl). Exemplaryphospholipids include phosphoglycerides such asdipalmitoylphosphatidylcholine, disteroylphosphatidylcholine,diarachidoylphosphatidylcholine, dibehenoylphosphatidylcholine,diphosphatidyl glycerol, short-chain phosphatidylcholines, long-chainsaturated phosphatidylethanolamines, long-chain saturatedphosphatidylserines, long-chain saturated phosphatidylglycerols, andlong-chain saturated phosphatidylinositols. Additional excipients aredisclosed in International Patent Publication No. WO 96/32149 and U.S.Pat. Nos. 6,358,530, 6,372,258 and 6,518,239.

In particular embodiments, the suspending particles may be formed usingone or more lipids, phospholipids or saccharides, as described herein.In some embodiments, suspending particles include one or moresurfactants. The use of suspending particles formed of or incorporatingone or more surfactants may promote absorption of the selected activeagent, thereby increasing bioavailability. The suspending particlesdescribed herein, such as, for example, suspending particles formedusing one or more lipids, can be formed to exhibit a desired surfacerugosity (roughness), which can further reduce inter-particleinteractions and improve aerosolization by reducing the surface areaavailable for particle-particle interaction. In further embodiments, ifsuitable, a lipid that is naturally occurring in the lung could be usedin forming the suspending particles, as such suspending particles thathave the potential to reduce opsonization (and thereby reducingphagocytosis by alveolar macrophages), thus providing a longer-livedcontrolled release particle in the lung.

In another aspect, the suspending particles utilized in the compositionsdescribed herein may be selected to increase storage stability of theselected active agent, similar to that disclosed in International PatentPublication No WO 2005/000267. For example, in one embodiment, thesuspending particles my include pharmaceutically acceptable glassstabilization excipients having a Tg of at least 55° C., at least 75°C., or at least 100° C. Glass formers suitable for use in compositionsdescribed herein include, but are not limited to, one or more oftrileucine, sodium citrate, sodium phosphate, ascorbic acid, inulin,cyclodextrin, polyvinyl pyrrolidone, mannitol, sucrose, trehalose,lactose, and, proline. Examples of additional glass-forming excipientsare disclosed in U.S. Pat. Nos. RE 37,872, 5,928,469, 6,258,341.

The suspending particles may be designed, sized and shaped as desired toprovide desirable stability and active agent delivery characteristics.In one exemplary embodiment, the suspending particles compriseperforated microstructures as described herein. Where perforatedmicrostructures are used as suspending particles in the compositionsdescribed herein, they may be formed using one or more excipients asdescribed herein. For example, in particular embodiments, perforatedmicrostructures may include at least one of the following: lipids,phospholipids, nonionic detergents, nonionic block copolymers, ionicsurfactants, biocompatible fluorinated surfactants and combinationsthereof, particularly those approved for pulmonary use. Specificsurfactants that may be used in the preparation of perforatedmicrostructures include poloxamer 188, poloxamer 407 and poloxamer 338.Other specific surfactants include oleic acid or its alkali salts. Inone embodiment, the perforated microstructures include greater thanabout 10% w/w surfactant.

In some embodiments, suspending particles may be prepared by forming anoil-in-water emulsion, using a fluorocarbon oil (e.g., perfluorooctylbromide, perfluorodecalin) which may be emulsified using a surfactantsuch as a long chain saturated phospholipid. The resultingperfluorocarbon in water emulsion may be then processed using a highpressure homogenizer to reduce the oil droplet size. The perfluorocarbonemulsion may be fed into a spray dryer, optionally with an active agentsolution, if it is desirable to include active agent within the matrixof the perforated microstructures. As is well known, spray drying is aone-step process that converts a liquid feed to a dried particulateform. Spray drying has been used to provide powdered pharmaceuticalmaterial for various administrative routes, including inhalation.Operating conditions of the spray dryer (such as inlet and outlettemperature, feed rate, atomization pressure, flow rate of the dryingair and nozzle configuration) can be adjusted to produce the desiredparticle size producing a yield of the resulting dry microstructures.Such methods of producing exemplary perforated microstructures aredisclosed in U.S. Pat. No. 6,309,623 to Weers et al.

Perforated microstructures as described herein may also be formedthrough lyophilization and subsequent milling or micronization.Lyophilization is a freeze-drying process in which water is sublimedfrom the composition after it is frozen. This process allows dryingwithout elevated temperatures. In yet further embodiments, thesuspending particles may be produced using a spray freeze dryingprocess, such as is disclosed in U.S. Pat. No. 5,727,333.

Furthermore, suspending particles as described herein may includebulking agents, such as polymeric particles. Polymeric polymers may beformed from biocompatible and/or biodegradable polymers, copolymers orblends. In one embodiment, polymers capable of forming aerodynamicallylight particles may be used, such as functionalized polyester graftcopolymers and biodegradable polyanhydrides. For example, bulk erodingpolymers based on polyesters including poly(hydroxy acids) can be used.Polyglycolic acid (PGA), polylactic acid (PLA) or copolymers thereof maybe used to form suspending particles. The polyester may include acharged or functionalizable group, such as an amino acid. For example,suspending particles may be formed of poly(D,L-lactic acid) and/orpoly(D,L-lactic-co-glycolic acid) (PLGA), which incorporate a surfactantsuch as DPPC.

Other potential polymer candidates for use in suspending particles mayinclude polyamides, polycarbonates, polyalkylenes such as polyethylene,polypropylene, poly(ethylene glycol), poly(ethylene oxide),poly(ethylene terephthalate), poly vinyl compounds such as polyvinylalcohols, polyvinyl ethers, and polyvinyl esters, polymers of acrylicand methacrylic acids, celluloses and other polysaccharides, andpeptides or proteins, or copolymers or blends thereof. Polymers may beselected with or modified to have the appropriate stability anddegradation rates in vivo for different controlled drug deliveryapplications.

The compositions described herein may include two or more species ofsuspending particles. Even further, compositions according to thepresent description can include suspending particles that includeglycopyrrolate incorporated into the suspending particles. Where activeagent is incorporated into suspending particles, the suspendingparticles will be of a respirable size and can be formulated andproduced using, for example, the methods and materials described herein.

Compositions formulated according to the present teachings can inhibitdegradation of active agent included therein. For example, in specificembodiments, the compositions described herein inhibit one or more offlocculation, aggregation and the solution mediated transformation ofactive agent material included in the compositions. The pharmaceuticalcompositions described herein are suited for respiratory delivery viaand MDI in a manner that achieves desirable delivered dose uniformity(“DDU”) of LABA and LAMA active agents, including potent and highlypotent LABA and LAMA agents throughout emptying of an MDI canister. Asis described in detail in the Examples included herein, even whendelivering very low doses of LAMA or LABA active agents, compositionsdescribed herein can achieve a DDU for the active agent of ±30%, orbetter throughout emptying of an MDI canister. In one such embodiment,compositions described herein achieve a DDU for the active agent of±25%, or better throughout emptying of an MDI canister. In yet anothersuch embodiment, compositions described herein achieve a DDU for theactive agent of ±20%, or better throughout emptying of an MDI canister.

Pharmaceutical compositions described herein also serve to substantiallypreserve FPF and FPD performance throughout emptying of an MDI canister,even after being subjected to accelerated degradation conditions. Forinstance, compositions according to the present description maintain asmuch as 80%, 90%, 95%, or more, of the original FPF and FPD performancethroughout emptying of an MDI canister, even after being subjected toaccelerated degradation conditions. Compositions described hereinprovide the added benefit of achieving such performance while beingformulated using non-CFC propellants. In specific embodiments, thecompositions described herein achieve desired one or all of a targetedDDU, FPF and FPD performance while being formulated with suspensionmedium including only one or more non-CFC propellants and without theneed to modify the characteristics of the non-CFC propellant, such as bythe addition of, for example, one or more cosolvent, antisolvent,solubilizing agent, adjuvant or other propellant modifying material.

In one embodiment, a co-suspension composition as described hereinincludes: a suspension medium comprising a pharmaceutically acceptableHFA propellant; a plurality of active agent particles comprisingglycopyrrolate, including any pharmaceutically acceptable salts, esters,isomers or solvates thereof, suspended in the suspension medium at aconcentration sufficient to provide a delivered dose of glycopyrrolateof between about 20 μg and about 150 μg per actuation of the metereddose inhaler; and a plurality of respirable suspending particlescomprising perforated microstructures as described herein exhibiting avolume median optical diameter of between about 1.5 μm and about 10 μm,wherein perforated microstructures associate with the plurality ofactive agent particles to form a co-suspension. In one such embodiment,the glycopyrrolate active agent particles are formed of crystallineglycopyrrolate material. In another such embodiment, the ratio of thetotal mass of the suspending particles to the total mass of the activeagent particles is selected from between about 3:1 and about 15:1 andbetween about 2:1 and 8:1. In yet another such embodiment, theglycopyrrolate active agent particles are formed of crystallineglycopyrrolate material and the ratio of the total mass of thesuspending particles to the total mass of the active agent particles isselected from between about 3:1 and about 15:1 and between about 2:1 and8:1. In still another such embodiment, the glycopyrrolate active agentparticles are formed of crystalline glycopyrrolate material, at least90% of the glycopyrrolate active agent particles by volume exhibit anoptical diameter of less than 7 μm, and the ratio of the total mass ofthe suspending particles to the total mass of the active agent particlesis selected from between about 3:1 and about 15:1 and between about 2:1and 8:1.

In another embodiment, a co-suspension composition as described hereinincludes: a suspension medium comprising a pharmaceutically acceptableHFA propellant; a plurality of active agent particles comprisingtiotropium, including any pharmaceutically acceptable salts, esters,isomers or solvates thereof, suspended in the suspension medium at aconcentration sufficient to provide a delivered dose of glycopyrrolateof between about 5 μg and about 40 μg per actuation of the metered doseinhaler; and a plurality of respirable suspending particles comprisingperforated microstructures as described herein exhibiting a volumemedian optical diameter of between about 1.5 μm and about 10 μm, whereinperforated microstructures associate with the plurality of active agentparticles to form a co-suspension. In one such embodiment, thetiotropium active agent particles are formed of crystalline tiotropiummaterial. In another such embodiment, the ratio of the total mass of thesuspending particles to the total mass of the active agent particles isselected from between about 3:1 and about 15:1 and between about 2:1 and8:1. In yet another such embodiment, the tiotropium active agentparticles are formed of crystalline tiotropium material and the ratio ofthe total mass of the suspending particles to the total mass of theactive agent particles is selected from between about 3:1 and about 15:1and between about 2:1 and 8:1. In still another such embodiment, thetiotropium active agent particles are formed of crystalline tiotropiummaterial, at least 90% of the tiotropium active agent particles byvolume exhibit an optical diameter of less than 7 μm, and the ratio ofthe total mass of the suspending particles to the total mass of theactive agent particles is selected from between about 3:1 and about 15:1and between about 2:1 and 8:1.

In another embodiment, a co-suspension composition as described hereinincludes: a suspension medium comprising a pharmaceutically acceptableHFA propellant; a plurality of active agent particles comprisingformoterol, including any pharmaceutically acceptable salts, esters,isomers or solvates thereof, suspended in the suspension medium at aconcentration sufficient to provide a delivered dose of formoterol ofbetween about 0.5 μg and about 10 μg per actuation of the metered doseinhaler; and a plurality of respirable suspending particles comprisingperforated microstructures as described herein exhibiting a volumemedian optical diameter of between about 1.5 μm and about 10 μm, whereinperforated microstructures associate with the plurality of active agentparticles to form a co-suspension. In one such embodiment, theformoterol active agent particles are formed of crystalline formoterolmaterial. In another such embodiment, the ratio of the total mass of thesuspending particles to the total mass of the active agent particles isselected from between about 3:1 and about 15:1 and between about 2:1 and8:1. In yet another such embodiment, the formoterol active agentparticles are formed of crystalline formoterol material and the ratio ofthe total mass of the suspending particles to the total mass of theactive agent particles is selected from between about 3:1 and about 15:1and between about 2:1 and 8:1. In still another such embodiment, theformoterol active agent particles are formed of crystalline formoterolmaterial, at least 90% of the formoterol active agent particles byvolume exhibit an optical diameter of less than 7 μm, and the ratio ofthe total mass of the suspending particles to the total mass of theactive agent particles is selected from between about 3:1 and about 15:1and between about 2:1 and 8:1.

In one embodiment, a co-suspension composition as described hereinincludes: a suspension medium comprising a pharmaceutically acceptableHFA propellant; a plurality of active agent particles comprisingformoterol, including any pharmaceutically acceptable salts, esters,isomers or solvates thereof, suspended in the suspension medium at aconcentration sufficient to provide a delivered dose of formoterol ofbetween about 2 μg and about 10 μg per actuation of the metered doseinhaler; and a plurality of respirable suspending particles comprisingperforated microstructures as described herein exhibiting a volumemedian optical diameter of between about 1.5 μm and about 10 μm, whereinperforated microstructures associate with the plurality of active agentparticles to form a co-suspension. In one such embodiment, theformoterol active agent particles are formed of crystalline formoterolmaterial. In another such embodiment, the ratio of the total mass of thesuspending particles to the total mass of the active agent particles isselected from between about 3:1 and about 15:1 and between about 2:1 and8:1. In yet another such embodiment, the formoterol active agentparticles are formed of crystalline formoterol material and the ratio ofthe total mass of the suspending particles to the total mass of theactive agent particles is selected from between about 3:1 and about 15:1and between about 2:1 and 8:1. In still another such embodiment, theformoterol active agent particles are formed of crystalline formoterolmaterial, at least 90% of the formoterol active agent particles byvolume exhibit an optical diameter of less than 7 μm, and the ratio ofthe total mass of the suspending particles to the total mass of theactive agent particles is selected from between about 3:1 and about 15:1and between about 2:1 and 8:1.

In another embodiment, a co-suspension composition as described hereinincludes: a suspension medium comprising a pharmaceutically acceptableHFA propellant; a plurality of active agent particles comprisingsalmeterol, including any pharmaceutically acceptable salts, esters,isomers or solvates thereof, suspended in the suspension medium at aconcentration sufficient to provide a delivered dose of salmeterol ofbetween about 8 μg and about 40 μg per actuation of the metered doseinhaler; and a plurality of respirable suspending particles comprisingperforated microstructures as described herein exhibiting a volumemedian optical diameter of between about 1.5 μm and about 10 μm, whereinperforated microstructures associate with the plurality of active agentparticles to form a co-suspension. In one such embodiment, thesalmeterol active agent particles are formed of crystalline salmeterolmaterial. In another such embodiment, the ratio of the total mass of thesuspending particles to the total mass of the active agent particles isselected from between about 3:1 and about 15:1 and between about 2:1 and8:1. In yet another such embodiment, the salmeterol active agentparticles are formed of crystalline salmeterol material and the ratio ofthe total mass of the suspending particles to the total mass of theactive agent particles is selected from between about 3:1 and about 15:1and between about 2:1 and 8:1. In still another such embodiment, thesalmeterol active agent particles are formed of crystalline salmeterolmaterial, at least 90% of the salmeterol active agent particles byvolume exhibit an optical diameter of less than 7 μm, and the ratio ofthe total mass of the suspending particles to the total mass of theactive agent particles is selected from between about 3:1 and about 15:1and between about 2:1 and 8:1.

III. METERED DOSE INHALER SYSTEMS

As described in relation to the methods provided herein, thecompositions disclosed herein may be used in an MDI system. MDIs areconfigured to deliver a specific amount of a medicament in aerosol form.In one embodiment, an MDI system includes a pressurized, liquid phaseformulation-filled canister disposed in an actuator formed with amouthpiece. The MDI system may include the formulations describedherein, which include a suspension medium, glycopyrrolate and at leastone species of suspending particles. The canister used in the MDI be anyof any suitable configuration, and in one exemplary embodiment, thecanister may have a volume ranging from about 5 mL to about 25 mL, suchas, for example a canister having a 19 mL volume. After shaking thedevice, the mouthpiece is inserted into a patient's mouth between thelips and teeth. The patient typically exhales deeply to empty the lungsand then takes a slow deep breath while actuating the cartridge.

Inside an exemplary cartridge is a metering valve including a meteringchamber capable of holding a defined volume of the formulation (e.g., 63μl or any other suitable volume available in commercially availablemetering valves), which is released into an expansion chamber at thedistal end of the valve stem when actuated. The actuator retains thecanister and may also include a port with an actuator nozzle forreceiving the valve stem of the metering valve. When actuated, thespecified volume of formulation travels to the expansion chamber, outthe actuator nozzle and into a high-velocity spray that is drawn intothe lungs of a patient.

IV. METHODS

Methods for formulating pharmaceutical compositions for respiratorydelivery of LAMA and LAMA active agents are provided herein. Inparticular embodiments, such methods involve the steps of providing asuspension medium, active agent particles selected from active agentparticles comprising a LAMA and active agent particles comprising aLABA, and one or more species of suspending particles, as describedherein, and combining such constituents to form a formulation whereinthe active agent particles associate with the suspending particles andco-locate with the suspending particles within the suspension mediumsuch that a co-suspension is formed. In one such embodiment, theassociation of the glycopyrrolate particles and the suspending particlesis such that they do not separate due to their different buoyancies in apropellant. As will be appreciated, the method may include providing twoor more species of suspending particles in combination with active agentparticles. In another embodiment, the method may include providing twoor more species of active agent particles and combining the two or morespecies of active agent particles with one or more species of suspendingparticles in a manner that results in a co-suspension. In certainembodiments, the active agent particles consist essentially of a LAMA orLABA active agent as described herein.

In specific embodiments of methods for providing a stabilizedcomposition of a LAMA or LABA active agent for pulmonary delivery, thepresent disclosure provides methods for inhibiting the solution mediatedtransformation of the LAMA or LABA active agent in a pharmaceuticalcomposition for pulmonary delivery. In one embodiment, a suspensionmedium as described herein, such as a suspension medium formed by an HFApropellant, is obtained. Suspending particles are also obtained orprepared as described herein. Active agent particles are also obtained,and the suspension medium, suspending particles and active agentparticles are combined to form a co-suspension wherein the active agentparticles associate with suspending particles and co-locate with thesuspending particles within the continuous phase formed by thesuspension medium. When compared to active agent particles contained inthe same suspension medium in the absence of suspending particles,co-suspensions according to the present description have been found toexhibit a higher tolerance to solution mediated phase transformationthat leads to irreversible crystal aggregation, and thus may lead toimproved stability and dosing uniformity.

In further embodiments, methods for forming stabilized compositions ofLAMA and LABA active agents for pulmonary delivery include forpreserving the FPF and/or FPD of the composition throughout emptying ofan MDI canister. In specific embodiments of methods for preserving theFPF and/or FPD provided by a pharmaceutical composition for pulmonarydelivery, a respirable co-suspension as described herein is providedwhich is capable of maintaining the FPD and/or the FPF to within ±20%,±10%, or even ±5% the initial FPD and/or FPF, respectively, throughoutemptying of an MDI canister. Such performance can be achieved even afterthe co-suspension is subjected to accelerated degradation conditions. Inone embodiment, a suspension medium as described herein, such as asuspension medium formed by an HFA propellant, is obtained. Suspendingparticles are also obtained or prepared as described herein. Activeagent particles are also obtained, and the suspension medium, suspendingparticles and active agent particles are combined to form aco-suspension wherein the glycopyrrolate particles associate withsuspending particles and co-locate with the suspending particles withinthe suspension medium. Even after exposure of such composition to one ormore temperature cycling events, the co-suspension maintains an FPD orFPF within ±20%, ±10%, or even ±5% of the respective values measuredprior to exposure of the composition to the one or more temperaturecycling events.

Methods for preparing an MDI for pulmonary delivery of LAMA or LABAactive agent are disclosed. The method of preparing the MDI may includeloading a canister, as described herein, with active agent particles andsuspending particles. An actuator valve can be attached to an end of thecanister and the canister sealed. The actuator valve may be adapted fordispensing a metered amount of the glycopyrrolate pharmaceuticalformulation per actuation. The canister can be charged with apharmaceutically acceptable suspension medium, such as a propellant asdescribed herein. Whereupon the active agent particles and suspendingparticles yield a stable co-suspension in the suspension medium.

In methods involving pulmonary delivery of a LAMA or LABA active agentusing compositions described herein, the compositions may be deliveredby an MDI. Therefore, in particular embodiments of such methods, an MDIloaded with a composition described herein is obtained, and a LAMA orLABA active agent is administered to a patient through pulmonarydelivery through actuation of the MDI. For example, in one embodiment,after shaking the MDI device, the mouthpiece is inserted into apatient's mouth between the lips and teeth. The patient typicallyexhales deeply to empty the lungs and then takes a slow deep breathwhile actuating the cartridge of the MDI. When actuated, the specifiedvolume of formulation travels to the expansion chamber, out the actuatornozzle and into a high-velocity spray that is drawn into the lungs of apatient. In one embodiment the dose of active agent delivered throughoutemptying of an MDI canister is not more than 30% greater than the meandelivered dose and is not less than 30% less than the mean delivereddose. Therefore, methods of achieving a desired DDU of glycopyrrolatedelivered from an MDI are also provided. In such embodiments, the methodmay include achieving a DDU for glycopyrrolate delivered from an MDIselected from, for example, a DDU of ±30%, or better, a DDU of ±25%, orbetter, and a DDU of ±20%, or better.

Methods for treating patients suffering from an inflammatory orobstructive pulmonary disease or condition are provided herein. Inspecific embodiments, such methods include pulmonary delivery of apharmaceutical composition described herein, and in certain suchembodiments, pulmonary administration of the pharmaceutical compositionis accomplished by delivering the composition using an MDI. The diseaseor condition to be treated can be selected from any inflammatory orobstructive pulmonary disease or condition that responds to theadministration of a LAMA or LABA agent. In particular embodiments, thepharmaceutical compositions described herein may be used in treating adisease or disorder selected from asthma, COPD, exacerbation of airwayshyper reactivity consequent to other drug therapy, allergic rhinitis,sinusitis, pulmonary vasoconstriction, inflammation, allergies, impededrespiration, respiratory distress syndrome, pulmonary hypertension,pulmonary vasoconstriction, emphysema, and any other respiratorydisease, condition, trait, genotype or phenotype that can respond to theadministration of a LAMA or LABA, alone or in combination with othertherapies. In certain embodiments, the pharmaceutical compositionsdescribed herein may be used in treating pulmonary inflammation andobstruction associated with cystic fibrosis.

Additionally, pharmaceutical compositions according to the presentdescription delivered from an MDI provide desirable pharmacodynamic (PD)performance. In particular embodiments, pulmonary delivery of thepharmaceutical compositions described herein results in rapid,significant improvement in the lung capacity, which can be characterizedby an improvement in the patient's forced expiratory volume in onesecond (FEV₁). For example, in particular embodiments, methods forachieving a clinically relevant increase in FEV₁ are provided, whereinsuch methods include providing a co-suspension composition comprising aLABA or LAMA active agent as described herein and administering suchcomposition to a patient experiencing pulmonary inflammation orobstruction via an MDI. For purposes of the present disclosure, aclinically relevant increase in FEV₁ is any increase of 100 ml orgreater, and in certain embodiments of the methods described herein,administration of compositions according to the present description topatient results in a clinically significant increase in FEV₁ within 1hour or less. In other such embodiments, methods for administering acomposition as described herein to a patient via an MDI result in aclinically significant increase in FEV1 within 0.5 hours or less. Thecompositions provided and delivered in such embodiments may include acomposition including a LAMA active agent or a composition including aLABA active agent as described herein.

In further embodiments, methods are provided for achieving an increasein FEV₁ greater than 100 ml. For example, in certain embodiments, themethods described herein include methods for achieving an FEV₁ of 150 mlor greater within a period of time selected from 0.5 hours or less, 1hour or less, and 1.5 hours or less. In other embodiments, the methodsdescribed herein include methods for achieving an FEV₁ of 200 ml orgreater within a period of time selected from 0.5 hours or less, 1 houror less, and 1.5 hours or less, and 2 hours or less. In certain suchembodiments, a composition comprising a LABA or LAMA active agent asdescribed herein is provided and administered to a patient experiencingpulmonary inflammation or obstruction via an MDI.

In still further embodiments, methods for achieving and maintaining aclinically significantly increase in FEV₁ are provided. In particularembodiments, upon administration of a single dose of a LABA or LAMAactive agent formulated in a composition as described herein to apatient via an MDI, a clinically significant increase in FEV₁ isachieved in a period of time selected from 0.5 hours or less, 1 hour orless, and 1.5 hours or less, and the clinically significant increase inFEV₁ is maintained for up 12 hours or more. In certain such embodiments,the increase in FEV₁ may be selected from an increase of 150 ml orgreater, 200 ml or greater and 250 ml or greater, and the increase inFEV₁ remains clinically significant for a time period selected from upto 4 hours, up to 6 hours, up to 8 hours, up to 10 hours, and up to 12hours, or more. In certain such embodiments, a composition comprising aLABA or LAMA active agent as described herein is provided andadministered a patient experiencing pulmonary inflammation orobstruction via an MDI.

Compositions, systems and methods described herein are not only suitedto achieving desirable pharmacodynamic performance in short periods oftime, but will achieve such results in a high percentage of patients.For example, methods are provided herein for achieving a 10% or greaterincrease in FEV₁ in 50% or more of patients experiencing pulmonaryinflammation or obstruction. For example, in particular embodiments,methods for achieving a 10% or greater increase in FEV₁ in a patientinclude providing a co-suspension composition comprising a LABA or LAMAactive agent as described herein and administering such composition viaan MDI to a patient experiencing pulmonary inflammation or obstruction.In certain such embodiments, administration of the composition resultsin 10% or greater increase in FEV₁ within a period of time selected from0.5 hours or less, 1 hour or less, 1.5 hours or less, and 2 hours in 50%or more of patients. In other such embodiments, administration of thecomposition results in 10% or greater increase in FEV₁ within a periodof time selected from 0.5 hours or less, 1 hour or less, 1.5 hours orless, and 2 or less hours in 60% or more of patients. In still othersuch embodiments, administration of the composition results in 10% orgreater increase in FEV₁ within a period of time selected from 0.5 hoursor less, 1 hour or less, 1.5 hours or less, and 2 hours or less in 70%or more of patients. In yet other such embodiments, administration ofthe composition results in 10% or greater increase in FEV₁ within aperiod of time selected from 0.5 hours or less, 1 hour or less, 1.5hours or less, and 2 or less hours in 80% or more of patients

In specific embodiments, the methods described herein facilitatetreatment of patients experiencing pulmonary inflammation orobstruction, wherein such methods include providing a co-suspensioncomposition comprising a LABA or LAMA active agent as described hereinand administering such composition to a patient experiencing pulmonaryinflammation or obstruction via an MDI and result in a high proportionof such patients experiencing either an increase from baseline in FEV₁of at least 200 ml or a 12%, or greater, increase from baseline in FEV₁coupled with total increase in FEV₁ of at least 150 ml. In certain suchembodiments, administration of the composition results in either anincrease from baseline in FEV₁ of at least 200 ml or a 12%, or greater,increase from baseline in FEV₁ coupled with total increase in FEV₁ of atleast 150 ml within a period of time selected from 1 hour or less, 1.5hours or less, 2 hours or less, and 2.5 hours or less in 50% or more ofpatients. In other such embodiments, administration of the compositionresults in an increase from baseline in FEV₁ of at least 200 ml or a12%, or greater, increase from baseline in FEV₁ coupled with totalincrease in FEV₁ of at least 150 ml within a period of time selectedfrom 1 hour or less, 1.5 hours or less, 2 hours or less, and 2.5 hoursor less in 60% or more of patients. In still other such embodiments,administration of the composition results in either an increase frombaseline in FEV₁ of at least 200 ml or a 12%, or greater, increase frombaseline in FEV₁ coupled with total increase in FEV₁ of at least 150 mlwithin a period of time selected from 1.5 hours or less, 2 hours orless, 2.5 hours or less, and 3 hours or less in 70% or more of patients.In yet other such embodiments, administration of the composition resultsin either an increase from baseline in FEV₁ of at least 200 ml or a 12%,or greater, increase from baseline in FEV₁ coupled with total increasein FEV₁ of at least 150 ml within a period of time selected from 1.5hours or less, 2 hours or less, 2.5 hours or less, and 3 hours or lessin 80% or more of patients.

In some embodiments, pharmaceutical compositions according to thepresent description delivered from an MDI provide improvement in thelung capacity, which can be characterized by an improvement inspiratorycapacity (IC), which is defined as the maximal volume of gas that can betaken into the lungs in a full inhalation following a normal expiration.For example, in particular embodiments, methods for achieving aclinically relevant increase in IC are provided, wherein such methodsinclude providing a co-suspension composition comprising a LABA or LAMAactive agent as described herein and administering such composition to apatient experiencing pulmonary inflammation or obstruction via an MDI.For purposes of the present disclosure, a clinically relevant increasein IC is any increase of 70 ml or greater, and in certain embodiments ofthe methods described herein, administration of compositions accordingto the present description to patient results in a clinicallysignificant increase in IC within 2 hours or less. In other suchembodiments, methods for administering a composition as described hereinto a patient via an MDI result in a clinically significant increase inIC within 1 hour or less. In other such embodiments, administration ofcompositions according to the present description to patient results inan increase in IC of 100 ml or greater within a period of time selectedfrom 1 hour or less and 2 hours or less. In still other suchembodiments, administration of compositions according to the presentdescription to patient results in an increase in IC of 150 ml or greaterwithin a period of time selected from 1 hour or less and 2 hours orless. In even further such embodiments, administration of compositionsaccording to the present description to patient results in an increasein IC of 300 ml or greater within a period of time selected from 1 houror less and 2 hours or less. The compositions provided and delivered insuch embodiments may include a composition including a LAMA active agentor a composition including a LABA active agent as described herein.

In particular embodiments of the methods described herein, thecompositions provided include a LAMA active agent. In such embodiments,the LAMA active agent can be selected from, for example, glycopyrrolate,dexipirronium, tiotropium, trospium, aclidinium, and darotropium,including any pharmaceutically acceptable salts, esters, isomers orsolvates thereof. In specific embodiments of the methods describedherein, the composition is a co-suspension composition as describedherein that includes glycopyrrolate or any pharmaceutically acceptablesalt, ester, isomer or solvate thereof. In other specific embodiments ofthe methods described herein, the composition is a co-suspensioncomposition as described herein that includes tiotropium or anypharmaceutically acceptable salt, ester, isomer or solvate thereof.Where glycopyrrolate or tiotropium is selected as the active agent foruse in the compositions produced or administered as part of the methodsdescribed herein, the amount of glycopyrrolate or tiotropium included inthe composition may be selected from, for example, those amountsspecifically disclosed with respect to the pharmaceutical compositionsdescribed herein.

In further embodiments of the methods described herein, the compositionsprovided include a LABA active agent. In such embodiments, the LABAactive agent can be selected from, for example, bambuterol, clenbuterol,formoterol, salmeterol, carmoterol, milveterol, indacaterol, andsaligenin- or indole-containing and adamantyl-derived β₂ agonists,including any pharmaceutically acceptable salts, esters, isomers orsolvates thereof. In specific embodiments of the methods describedherein, the composition is a co-suspension composition as describedherein that includes formoterol or any pharmaceutically acceptable salt,ester, isomer or solvate thereof. In other specific embodiments of themethods described herein, the composition is a co-suspension compositionas described herein that includes salmeterol or any pharmaceuticallyacceptable salt, ester, isomer or solvate thereof. Where formoterol orsalmeterol is selected as the active agent for use in the compositionsproduced or administered as part of the methods described herein, theamount of formoterol or salmterol included in the composition may beselected from, for example, those amounts specifically disclosed withrespect to the pharmaceutical compositions described herein.

Compositions, methods and systems described herein provide desirabledose efficiency and dose response for LAMA or LABA active agentsformulated for pulmonary delivery. For example, pulmonary delivery ofglycopyrrolate for treatment of conditions such as COPD has beenpreviously suggested or reported by Schroeckenstein et al., J. AllergyClin. Immunol., 1988; 82(1): 115-119, Leckie et al., Exp. Opin. Invest.Drugs, 2000; 9(1): 3-23, Skorodin, Arch. Intern. Med., 1993; 153:814-828, Walker et al., Chest, 1987; 91(1): 49-51, and InternationalPatent Publication WO/1997/039758. These references report a minimumeffective dose for glycopyrrolate of 200 μg-1,000 μg. Such dosingrequirements are in line with human clinical results reported byBannister et al. in U.S. Pat. No. 7,229,607, wherein subjects were givena 480 μg dose of glycopyrrolate. As is described in Example 6 providedherein, compositions of glycopyrrolate prepared according to the presentdescription and delivered to human subjects via an MDI achieved quickonset of action and clinically relevant improvements in FEV₁ and IC inaccordance with the methods detailed herein, even when deliveringsignificantly smaller doses of glycopyrrolate (the largest single dosedelivered in the study was 144 μg).

Singh et al. [D Singh, P A Corris, and S D Snape. “NVA237, a once-dailyinhaled antimuscarinic, provides 24-hour bronchodilator efficacy inpatients with moderate to-severe COPD” Poster presented at the AmericanThoracic Society International Conference, San Diego, Calif., May 19-24,2006] reported clinical work wherein glycopyrrolate was administered tohuman subjects via pulmonary delivery at doses of 20 μg, 125 μg, 250 μg,and 400 μg. Though such doses ranged below the 200 μg thresholdpreviously reported, as is also detailed in Example 6, compositions ofglycopyrrolate formulated and delivered as described herein stillachieved a relatively improved dose efficiency. For example, changes inFEV₁ AUC achieved by glycopyrrolate co-suspensions as described andevaluated in the clinical trial described in Example 6 are compared tothose achieved by the compositions of Singh et al. in FIG. 10. The 18 μgglycopyrrolate dose from Example 6 provided significantly betterbronchodilator response than the 20 μg dose reported by Singh et al.,and the 36 μg and 144 μg glycopyrrolate doses from Example 6 providingcomparable bronchodilator response to the 125 μg and 250 μg doses,respectively, reported by Singh et al.

In particular embodiments, methods for achieving desired pharmacodynamiceffects are provided, wherein the methods include administering aco-suspension composition as described herein to a patient via a metereddose inhaler, wherein the co-suspension includes glycopyrrolate activeagent particles as described herein to a patient via a metered doseinhaler such that a delivered dose of no more than 150 μg glycopyrrolateis administered to the patient. In one embodiment, a method forachieving a clinically significant increase in FEV₁ is provided, whereinthe method includes administering a co-suspension as described hereincomprising glycopyrrolate active agent particles to a patient via ametered dose inhaler such that a delivered dose of no more than 150 μgglycopyrrolate is administered to the patient. In one such embodiment, adelivered dose of no more than 100 μg glycopyrrolate is administered tothe patient, and in another embodiment, a delivered dose of no more than80 μg glycopyrrolate is administered to the patient. Even where doses ofno more than 80 μg, no more than 100 μg glycopyrrolate, or no more than150 μg glycopyrrolate are administered to the patient, in particularembodiments, the clinically significant increase in FEV1 is achieved in1 hour or less. In some such embodiments, the clinically significantincrease in FEV1 is achieved in 0.5 hours or less.

In further embodiments, methods are provided for achieving an increasein FEV₁ greater than 100 ml, wherein the methods include administering aco-suspension as described herein comprising glycopyrrolate active agentparticles to a patient via a metered dose inhaler such that a delivereddose of no more than 150 μg glycopyrrolate is administered to thepatient. For example, in certain embodiments, methods for achieving anFEV₁ of 150 ml or greater within a period of time selected from 0.5hours or less, 1 hour or less, and 1.5 hours or less, are provided,wherein the methods include administering a co-suspension as describedherein comprising glycopyrrolate active agent particles to a patient viaa metered dose inhaler such that a delivered dose of no more than 150 μgglycopyrrolate is administered to the patient. In other embodiments, themethods described herein include methods for achieving an FEV₁ of 200 mlor greater within a period of time selected from 0.5 hours or less, 1hour or less, and 1.5 hours or less, and 2 hours or less, wherein themethods include administering a co-suspension as described hereincomprising glycopyrrolate active agent particles to a patient via ametered dose inhaler such that a delivered dose of no more than 150 μgglycopyrrolate is administered to the patient.

In still further embodiments, methods for achieving and maintaining aclinically significantly increase in FEV₁ are provided, wherein themethods include administering a co-suspension as described hereincomprising glycopyrrolate active agent particles to a patient via ametered dose inhaler such that a delivered dose of no more than 150 μgglycopyrrolate is administered to the patient. In certain suchembodiments, upon administration of a single delivered dose ofglycopyrrolate of no more than 150 μg, a clinically significant increasein FEV₁ is achieved in a period of time selected from 0.5 hours or less,1 hour or less, and 1.5 hours or less, and the clinically significantincrease in FEV₁ is maintained for up 12 hours or more. For example, inparticular embodiments, the increase in FEV₁ may be selected from anincrease of 150 ml or greater, 200 ml or greater and 250 ml or greater,and the increase in FEV₁ remains clinically significant for a timeperiod selected from up to 4 hours, up to 6 hours, up to 8 hours, up to10 hours, and up to 12 hours, or more.

Methods for achieving an increase from baseline in FEV₁ of at least 200ml or a 12%, or greater, increase from baseline in FEV₁ coupled withtotal increase in FEV₁ of at least 150 ml are also provided, wherein themethods include administering a co-suspension as described hereincomprising glycopyrrolate active agent particles to a patient via ametered dose inhaler such that a delivered dose of no more than 150 μgglycopyrrolate is administered to the patient. In certain suchembodiments, administration of a delivered dose of no more than 150 μgglycopyrrolate from a co-suspension as described herein via a metereddose inhaler results in either an increase from baseline in FEV₁ of atleast 200 ml or a 12%, or greater, increase from baseline in FEV₁coupled with total increase in FEV₁ of at least 150 ml within a periodof time selected from 1 hour or less, 1.5 hours or less, 2 hours orless, and 2.5 hours or less in 50% or more of patients. In other suchembodiments, administration of a delivered dose of no more than 150 μgglycopyrrolate from a co-suspension as described herein via a metereddose inhaler results in an increase from baseline in FEV₁ of at least200 ml or a 12%, or greater, increase from baseline in FEV₁ coupled withtotal increase in FEV₁ of at least 150 ml within a period of timeselected from 1 hour or less, 1.5 hours or less, 2 hours or less, and2.5 hours or less in 60% or more of patients. In still other suchembodiments, administration of a delivered dose of no more than 150 μgglycopyrrolate from a co-suspension as described herein via a metereddose inhaler results in either an increase from baseline in FEV₁ of atleast 200 ml or a 12%, or greater, increase from baseline in FEV₁coupled with total increase in FEV₁ of at least 150 ml within a periodof time selected from 1.5 hours or less, 2 hours or less, 2.5 hours orless, and 3 hours or less in 70% or more of patients. In yet other suchembodiments, administration of a delivered dose of no more than 150 μgglycopyrrolate from a co-suspension as described herein via a metereddose inhaler results in either an increase from baseline in FEV₁ of atleast 200 ml or a 12%, or greater, increase from baseline in FEV₁coupled with total increase in FEV₁ of at least 150 ml within a periodof time selected from 1.5 hours or less, 2 hours or less, 2.5 hours orless, and 3 hours or less in 80% or more of patients.

Methods for achieving a clinically significant increase in IC areprovided, wherein the methods include administering a co-suspension asdescribed herein comprising glycopyrrolate active agent particles to apatient via a metered dose inhaler such that a delivered dose of no morethan 150 μg glycopyrrolate is administered to the patient. In certainsuch embodiments, administration of a delivered dose of no more than 150μg glycopyrrolate from a co-suspension as described herein via a metereddose inhaler results in a clinically significant increase in IC within 1hour or less. In other such embodiments, administration of a delivereddose of no more than 150 μg glycopyrrolate from a co-suspension asdescribed herein via a metered dose inhaler results in an increase in ICof 100 ml or greater within a period of time selected from 1 hour orless and 2 hours or less. In still other such embodiments,administration of a delivered dose of no more than 150 μg glycopyrrolatefrom a co-suspension as described herein via a metered dose inhalerresults in an increase in IC of 150 ml or greater within a period oftime selected from 1 hour or less and 2 hours or less. In even furthersuch embodiments, administration of a delivered dose of no more than 150μg glycopyrrolate from a co-suspension as described herein via a metereddose inhaler results in an increase in IC of 300 ml or greater within aperiod of time selected from 1 hour or less and 2 hours or less.

The specific examples included herein are for illustrative purposes onlyand are not to be considered as limiting to this disclosure. Moreover,the compositions, systems and methods disclosed herein have beendescribed in relation to certain embodiments thereof, and many detailshave been set forth for purposes of illustration, it will be apparent tothose skilled in the art that the invention is susceptible to additionalembodiments and that certain of the details described herein may bevaried without departing from the basic principles of the invention. Anyactive agents and reagents used in the following examples are eithercommercially available or can be prepared according to standardliterature procedures by those skilled in the art of organic synthesis.The entire contents of all publications, patents, and patentapplications referenced herein are hereby incorporated herein byreference.

Example 1

Active agent particles formed of glycopyrrolate (Pyrrolidinium,3-((cyclopentylhydroxyphenylacetyl)oxy)-1,1-dimethyl-, bromide) wereformed by micronizing glycopyrrolate using a jet mill. The particle sizedistribution of the micronized glycopyrrolate (GP) was determined bylaser diffraction. 50% by volume of the micronized particles exhibitedan optical diameter smaller than 2.1 μm, 90% by volume were smaller than5 μm.

Suspending particles were manufactured as follows: 500 mL of afluorocarbon-in water emulsion of PFOB (perfluoroctyl bromide)stabilized by a phospholipid was prepared. 18.7 g of the phospholipid,DSPC (1,2-Distearoyl-sn-Glycero-3-Phosphocholine), and 1.3 g of calciumchloride were homogenized in 400 mL of hot water (75° C.) using a highshear mixer. 100 mL of PFOB were added slowly during homogenization. Theresulting coarse emulsion was then further homogenized using a highpressure homogenizer (Model C3, Avestin, Ottawa, Calif.) at pressures ofup to 170 MPa for 5 passes.

The emulsion was spray dried in nitrogen using the following spraydrying conditions: Inlet temperature 95° C., outlet temperature 72° C.,emulsion feed rate 2.4 mL/min, total gas flow 525 L/min. The particlesize distribution of the suspending particles was determined by laserdiffraction. 50% by volume of the suspending particles were smaller than2.9 μm, the Geometric Standard Deviation of the distribution was 1.8.

Metered dose inhalers were prepared by weighing the target masses ofmicronized GP particles and suspending particles into fluorinatedethylene polymer (FEP) coated aluminum canisters (Presspart, Blackburn,UK) with 19 mL volume. The target masses and the target delivered doseassuming 20% actuator deposition are given in Table 1 for five differentconfigurations (configurations 1A through 10 representing differentsuspensions of GP particles and suspending particles; configuration 1Drepresenting GP particles alone; configuration 1E representingsuspending particles alone). The canisters were crimp sealed with 63 μlvalves (# BK 357, Bespak, King's Lynn, UK) and filled with 12.4 g of HFA134a (1,1,1,2-tetrafluoroethane) (Ineos Fluor, Lyndhurst, UK) byoverpressure through the valve stem. After injecting the propellant, thecanisters were sonicated for 15 seconds and agitated on a wrist actionshaker for 30 minutes. The canisters were fitted with polypropyleneactuators with a 0.3 mm orifice (# BK 636, Bespak, King's Lynn, UK).Additional inhalers for visual observation of suspension quality wereprepared using glass vials.

TABLE 1 Results for Glycopyrrolate Co-suspensions of Example 1 TargetConfig- GP Suspending delivered Delivered uration (mg/ particles doseDose FPF MMAD ID can) (mg/can) (μg) (μg) (%) (μm) 1A 3.4 61 16.5 17.841.3 3.7 1B 4.1 61 20 19.4 42.0 3.9 1C 4.1 15 20 19.2 42.7 3.2 1D 4.1 020 11.1-15.3 27.0 3.3 1E 0 61 — —  53.6 * 3.2 * Based on DSPC assay.

Aerosol performance was assessed shortly after manufacturing inaccordance with USP <601> (United States Pharmacopeia Monograph 601). ANext Generation Impactor (NGI) operated at a flow rate of 30 L/min wasused for determination of particle size distribution. Sample canisterswere seated into an actuator with two waste actuations and twoadditional waste priming actuations. Five actuations were collected inthe NGI with a USP throat attached. The valve, actuator, throat, NGIcups, stages, and filter were rinsed with volumetrically dispensedsolvent. The sample solutions were assayed using a drug specificchromatographic method. The fine particle fraction was defined using thesum of stages 3 through filter. Delivered dose uniformity through usetesting was performed using a Dose Uniformity Sampling Apparatus asdescribed in USP <601>. Inhalers were seated and primed as describedbefore. Two actuations were collected and assayed at beginning, middleand end of use.

Visual observation of the co-suspended configurations (1A, 1B, 1C)showed no sedimentation of drug crystals. The suspension flocculatedslowly and formed a homogeneous, single cream layer similar to thecomparator configuration 1E, which included suspending particlessuspended alone. In contrast, the micronized GP particles alone(configuration 1D) flocculated and sedimented quickly. Configuration 1Bshowed no indication of separation of GP particles from the suspendingparticles even after centrifugation at 35 g for 20 minutes. The sameresult was observed (i.e., lack of GP particle separation) whencentrifuged up to 200 g. Configuration 1C (low suspending concentration)showed a small amount of GP crystals settling out after centrifugationat 35 g for 20 minutes.

While the co-suspended configurations achieved a delivered dose within10% of target, the GP particles suspended alone showed much highervariability in delivered dose in a range significantly below target. Thefine particle fraction relative to configuration 1D was improved by morethan 50%. The MMADs of the co-suspended configurations were acceptableand depended on the suspension concentration of the suspendingparticles. The delivered dose uniformity through use was tested forconfigurations 1B and 1C. All individual delivered doses were within±20% of mean. The results showed that the drug crystals forming the GPparticles associate to the suspending particles, a co-suspension wasformed, and the aerosol performance of the co-suspension was mostlydetermined by the suspending particles.

The association between GP crystals and suspending particles was strongenough to overcome buoyancy forces, as it was observed that GP crystalsdo not separate from the perforated microstructures and settling of thecrystals is inhibited.

Example 2

Glycopyrrolate (GP) particles were formed by micronization using a jetmill. Suspending particles were manufactured as described in Example 1.The particle size distribution of the micronized GP was determined bylaser diffraction. 50% by volume of the micronized particles exhibitedan optical diameter smaller than 1.7 μm, 90% by volume exhibited anoptical diameter smaller than 4.1 μm. Five different lots of metereddose inhalers were different lots were made. For configurations 2A, 2Band 2C the total concentration of DSPC, CaCl₂, and GP in the feedstockwas 40 mg/mL, for configuration 2D and 2E this concentration wasdoubled.

Metered dose inhalers were prepared by weighing the target masses of GPparticles and suspending particles into canisters as described inExample 1. No further excipients were used. The target masses were 4mg/canister for GP particles and 60 mg/canister for the suspendingparticles, resulting in a suspending particle to GP particle ratio of 15for configurations 2A and 2D. The target masses were 5.1 mg/canister forGP particles and 51 mg/canister for the suspending particles, resultingin a suspending particle to GP particle ratio of 10 for configuration2B. The target masses were 8 mg/canister for GP particles and 60mg/canister for the suspending particles, resulting in a suspendingparticle to GP particle ratio of 7.5 for configurations 2C and 2E.Propellant and container closure system were as described in Example 1.

The GP crystals were placed in HFA 134a in a canister under pressure andwere equilibrated for 3 weeks at room temperature to determine theirsolubility in the propellant. The samples were filtered under pressureat ambient temperature through filters with a pore width of 0.22 μm. Thefiltrate was evaporated and the GP dissolved in methanol andchromatographically analyzed. A solubility of 0.17±0.07 μg/g was found.Using this value it was determined that 2.1 μg or 0.05% of GP present inthe canister dissolved in the propellant. Previous articles teach thatmicrocrystalline material with a measurable solubility in the propellantwill not be physically stable due to solution mediated transformation[N. C. Miller, The Effects of Water in Inhalation Suspension AerosolFormulations, in: P. A. Byron, Ed., Respiratory Drug Delivery, CRCPress, 1990, p 250], or that actives with solubility's above 0.1 μg/gshould be formulated with an adjuvant to prevent a solution mediatedtransformation [P. Rogueda, Novel Hydrofluoroalkane SuspensionFormulations for Respiratory Drug Delivery, Expert Opin. Drug Deliv. 2,625-638, 2005].

The filled metered dose inhalers were stored valve down without overwrapat two different conditions: 1) refrigerated at 5° C.; and 2) roomtemperature at 25° C./60% RH. Aerosol performance and delivered doseuniformity tests as described in Example 1 were carried out at differenttime points. The results, which are summarized in Table 2, show a stablefine particle fraction at refrigerated and room temperature conditions.

TABLE 2 Fine particle fraction of configurations in Example 2 FPF in % #Storage Initial 2 months 3 months 6 months 2A 5° C. 49 51 52 — 25°C./60% RH 48 51 — 2B 25° C./60% RH 50 46 49 48 2D 5° C. 51 54 54 — 25°C./60% RH 46 49 49

Configurations 2C and 2E were subjected to a temperature cycling test.The canisters were subjected to −5° C. and 40° C. alternating betweentemperatures every 6 hours for a total duration of twelve weeks. Fineparticle fraction was 53% for both configurations at the beginning ofthe study. After twelve weeks of cycling the FPF was unchanged, i.e. at55% for configuration 2C and at 53% for configuration 2E.

The delivered dose uniformity through use was tested at the 1, 2 and 6month time points. All individual delivered doses were within ±20% ofmean. FIGS. 1 and 2 show the aerosol particle size distributions asmeasured by the NGI for configurations 2A and 2B, respectively. Alsoshown are the amounts of drug recovered from actuator, and from theinduction port (throat) and its mouth piece adaptor. Recovered massesare expressed as percent of nominal dose. For configuration 2A,aerodynamic particle size distribution individual replicates are shownat 4, 8 and 12 weeks and at 8, 12 and 24 week for configuration 2B.Though there is a measurable fraction of the suspended GP dissolved inthe propellant, there is no evidence of a coarsening of the sizedistributions. Moreover, as evidenced by these Examples, the aerosolperformance of a co-suspension at suitable suspending particle to GPratios is determined largely by the suspending particles.

Example 3

Several similar batches of suspending particles were made as describedin Example 1. The suspending particles were combined with glycopyrrolate(GP) particles that were micronized to different extents, using twodifferent types of jet mills with various milling parameters. Theoptical diameter and particle size distribution of the micronized GPparticles was determined by laser diffraction. Table 3 lists the d₅₀ andd₉₀ values for the different lots of micronized material used. d₅₀ andd₉₀ denote the particle size at which the cumulative volume distributionreported by the particle sizing instrument reaches 50% and 90%respectively.

Twelve different lots of metered dose inhalers were prepared asdescribed in Example 1. In all cases the suspension concentration of GPparticles in HFA 134a was in the range of 0.32-0.45 mg/mL and thesuspension concentration of the suspending particles was in the range of5.8-6.1 mg/mL. The configurations were deemed similar enough to pool thedata for a meta-analysis presented in this Example.

The filled metered dose inhalers were stored valve down without overwrapat two different conditions: refrigerated at 5° C. and controlled roomtemperature at 25° C./60% RH. Aerosol performance tests as described inExample 1 were carried out at different time points. The results did notshow any statistically significant trend as a function of time up totwelve weeks of storage. No difference between room temperature storageand refrigerated storage was discernible. Hence, results from differentstress conditions and time points were pooled to determine how theparticle size distribution of the micronized material affects aerosolperformance.

Table 3 summarizes the MMAD results of the meta-analysis. The firstcolumn describes the six different configurations. The second columnidentifies how many individual lots were used in the compilation of thedata for the respective configuration. The third column lists the numberof individual MMAD determinations used to calculate the average MMAD forthe respective configuration. Columns four and five show the d₉₀ and d₅₀of the micronized material used to manufacture the co-suspensions. Theresults are sorted by d₉₀ value from coarse to fine. The last twocolumns display the average MMAD and standard deviation.

TABLE 3 Pooled MMAD results for 12 glycopyrrolate co-suspensions, sortedby the d₉₀ of the micronized glycopyrrolate particles. Number of AverageLot number MMAD d₉₀ d₅₀ MMAD ID of lots measurements (μm) (μm) (μm) SD3A 3 21 5.0 1.8 4.0 0.28 3B 2 9 4.9 2.1 4.1 0.37 3C 1 6 4.8 1.8 3.6 0.123D 1 4 4.3 1.7 3.5 0.22 3E 3 20 4.1 1.6 3.7 0.28 3F 2 10 3.5 1.7 3.60.10

These results show a weak dependence of MMAD on the d₉₀ of themicronized material. A similar analysis for the d₅₀ showed nostatistically significant trend. It can be concluded that changes in thesize distribution of the micronized material (e.g., different micronizedmaterial lots, or induced by solution mediated transformations) lead toonly minor differences in the size distribution of the aerosol emittedfrom the metered dose inhaler.

Example 4

Micronized glycopyrrolate (GP) particles were formed tested as describedin Example 1. The optical diameter of the micronized GP particles wasdetermined and 50% by volume of the micronized GP particles were smallerthan 1.7 μm, 90% by volume were smaller than 3.8 μm.

Five batches of suspending particles were made as described inExample 1. The batches differed in concentration, C_(F), and volumefraction of PFOB, V_(PFOB), of the feed emulsion prior to spray drying,ranging from 20 mg/mL to 160 mg/mL and 20% to 40%, respectively. Thedifferent configurations are described in Table 4.

Metered dose inhalers were prepared by weighing the target masses ofmicronized GP and suspending particles into coated glass vials with 15mL volume. The target suspension concentrations and suspending particleto GP ratios are given in Table 4 for the 26 different vials tested. Thecanisters were crimp sealed with 63 μl valves (Valois, Les Vaudreuil,France) and filled with 10 g or 12 g of HFA 134a(1,1,1,2-tetrafluoroethane) (Ineos Fluor, Lyndhurst, UK) by overpressurethrough the valve stem. After injecting the propellant, the canisterswere sonicated for 15 seconds and agitated on a wrist action shaker for30 minutes.

As described in Example 1, micronized GP particles formulated aloneflocculated and sedimented quickly. The glass vials in this example wereleft to settle for at least 24 h without agitation and then it wastested by visual observation whether the crystal, GP particles wereco-suspended completely. For the vials marked with “Yes” in Table 4, noGP particles were observed at the bottom of the vials, except for veryfew foreign particulates in some vials. Occasional foreign particleswere also visible in a similar very low amount in vials filled withsuspending particles only. For the vials marked “Partial,” a fraction ofthe GP particles was visible at the bottom of the vial.

TABLE 4 Co-suspension observations for glycopyrrolate configurationswith various suspending particle to glycopyrrolate particle ratios.Suspending C_(s) (mg/mL) particle to C_(F) in Suspending glycopyrrolateCo- # mg/mL V_(PFOB) (%) particle particle ratio suspension 4A 20 40 1.83.8 Partial 20 40 7.2 15 Yes 4B 40 40 3.0 1.9 Partial 40 40 1.8 3.8Partial 40 40 3.0 3.8 Yes 40 40 6.0 3.8 Yes 40 40 9.0 5.6 Yes 40 40 3.07.5 Yes 40 40 6.0 7.5 Yes 40 40 9.0 11.3 Yes 40 40 6.0 15 Yes 40 40 7.215 Yes 40 40 9.0 22.5 Yes 4C 80 20 3.0 1.9 Partial 80 20 3.0 3.8 Partial80 20 6.0 3.8 Yes 80 20 9.0 5.6 Yes 80 20 3.0 7.5 Yes 80 20 6.0 7.5 Yes80 20 9.0 11.3 Yes 80 20 6.0 15 Yes 80 20 9.0 22.5 Yes 4D 80 40 1.8 3.8Partial 80 40 7.2 15 Yes 4E 160 40 1.8 3.8 Partial 160 40 7.2 15 Yes

Example 5

Glycopyrrolate (GP) particles were micronized with a jetmill and testedas described in Example 1. 50% by volume of the micronized particlesexhibited an optical diameter smaller than 1.7 μm, 90% by volumeexhibited an optical diameter smaller than 4.4 μm.

Six batches of suspending particles were made by spray drying asdescribed in Example 1. Configuration 5A was spray dried from anemulsion. Configuration 5B was manufactured in a similar fashion butusing dipalmitoylphosphatidylcholine (DPPC) instead of DSPC.Configuration 5C was spray dried from an ethanolic solution. Forconfigurations 5D, 5E, and 5F, saccharides were spray dried from aqueoussolution. The spray drying parameters for all configurations are givenin Table 5a.

TABLE 5a Suspending particle configurations used in Example 5. SprayDrying Parameters Feed Total Powder Feed C_(F) rate Gas Lot compositioncomposition (mg/ (mL/ T_(in) T_(out) Flow # (% w/w) (% v/v) mL) min) (°C.) (° C.) (L/min) 5A 93.5% DSPC 80% H₂0 40 2.4 95 72 526 6.5% CaCl₂ 20%PFOB 5B 92.9% DPPC 70% H₂0 60 2.4 95 67 525 7.1% CaCl₂ 30% PFOB 5C 100%DSPC 95% Ethanol 100 5 95 70 520 5% PFOB 5D 100% Lactose 100% H₂0 100 495 70 668 5E 100% 100% H₂0 10 2.4 100 68 527 Trehalose 5F 100% 100% H₂089 4 100 71 670 Trehalose

The particle size distribution of the suspending particles wasdetermined by laser diffraction. The volume median optical diameter,VMD, and geometric standard deviation, GSD, for the differentconfigurations are given in Table 5b.

TABLE 5b Characteristics of suspending particle configurations used inExample 5. VMD Co- Lot # (μm) GSD Separation suspension Comment 5A 3.61.8 Creams Yes No or few 5B 3.6 1.8 Creams Yes crystals visible 5C 1.21.9 Creams Partial on bottom of vials 5D 1.7 2.3 Sediments Yes Causes GP5E 0.9 1.7 Sediments Yes crystals to 5F 1.7 2.4 Sediments Yes sedimentwith the suspending particles

Electron micrographs of the suspending particles showed a variety ofmorphologies, summarized in FIG. 3. The particles that were spray driedfrom emulsion, 5A and 5B, had high porosity and low density. The DSPCparticle spray dried from an ethanolic solution, 5C, showed a muchsmaller particle size with no noticeable porosity, indicating a highdensity. All saccharides produced smooth particles with no visibleporosity. Configuration 5E had the smallest particles, as expected dueto its low feed concentration.

Metered dose inhalers were prepared by weighing the 4 mg of micronizedGP particles and 60 mg of suspending particles into coated glass vialswith 15 mL volume. The canisters were crimp sealed with 63 μl valves(Valois DF30/63 RCU, Les Vaudreuil, France) and filled with 9.5 mL ofHFA 134a (Ineos Fluor, Lyndhurst, UK) by overpressure through the valvestem. After injecting the propellant, the canisters were sonicated for15 seconds and agitated on a wrist action shaker for 30 minutes.Additional inhalers with suspending particles only were manufactured ascontrol for each configuration.

The suspending particles in Examples 5A, 5B, and 5C, have true densitieslower than the propellant. They formed a cream layer and were tested forthe presence of a co-suspension as described in Example 4. No GPparticles were visible at the bottom of the vials for configuration 5Aand 5B. Configuration 5C formed a partial co-suspension.

The saccharide particles sediment because they have a higher truedensity than the propellant. However, all control vials for thesaccharide configurations showed a significantly faster sedimentationrate than micronized GP particles alone. In configurations 5D, 5E, and5F, the sedimentation rate was similar to that of the control vials withthe suspending particles alone and faster than the micronized GPparticles alone, demonstrating the association of the GP crystals withthe suspending particles. A co-suspension was formed in these cases.FIG. 4 shows an example of this behavior for configuration 5D. The glassvial was observed one minute after agitation. The co-suspension hasalready settled leaving a clear propellant layer, while in the controlcontaining GP particles alone, most of the crystals are still suspendedin the propellant.

Example 6

Pharmaceutical compositions according to the present description wereevaluated in a multi-center clinical trial. MDI devices containing apharmaceutical composition of glycopyrrolate prepared according to thepresent description were provided.

Suspending particles used were prepared in a similar manner described inExample 1. MDI manufacturing was accomplished using a drug additionvessel (DVA) by first adding half of Suspending particle quantity, nextfilling the microcrystalline GP, and lastly adding the remaining half ofsuspending particles to the top. Materials were added to the vessel in ahumidity controlled environment of <10% RH. The DAV was then connectedto a 4 L suspension vessel and flushed with HFA 134a propellant and thenmixed. The temperature inside the vessel was maintained at 21-23° C.throughout the entire batch production. After recirculation of the batchfor 30 min canisters were filled with the suspension mixture through 50μL EPDM valves. Sample canisters were then selected at random for totalcanister assay to ensure correct formulation quantities. The freshlymanufactured co-suspension MDI batch was then placed on one weekquarantine before initial product performance analysis.

The composition was formulated and the MDI devices configured to providea dose of 18 μg glycopyrrolate per MDI actuation.

The study was a randomized, double-blind, four-period, six-treatment,placebo and active-controlled crossover study which evaluated singleadministration of 4 ascending doses of glycopyrrolate in patients withmild to moderate COPD compared to placebo and open label tiotropium (18μg via the Spiriva Handihaler) as an active control. The six studytreatments were Glycopyrrolate MDI at doses of 18, 36, 72 and 144 μgwere achieved by one, two, four or eight consecutive actuations of the18 μg per actuation Glycopyrrolate MDI. Tiotropium Handihaler at 18 μg,and Placebo MDI, which was identical to the Glycopyrrolate MDI butwithout glycopyrrolate. Each patient was randomized to one of sixpossible sequences that included four of the study treatments. Eachsequence included two or three Glycopyrrolate MDI doses, which wereadministered in ascending order to each patient. Glycopyrrolate MDI andPlacebo MDI treatments were blinded and tiotropium was open label.Thirty-three patients were enrolled and analyzed for safety; thirtypatients were analyzed for efficacy. Peak improvement in FEV₁ relativeto test day baseline (FEV₁ is the maximum volume of air exhaled duringthe first second of maximum effort from a maximum inhalation), time toonset of action, time to peak FEV₁, FEV₁ AUC₀₋₁₂, FEV₁ AUC₀₋₂₄, FEV₁AUC₁₂₋₂₄, 12 and 24-hour trough FEV₁, and similar analyses for peakexpirator flow rate (PEFR) and FVC, as well as peak improvement ininspiratory capacity (IC) were evaluated. Blood samples were collectedpre-dose and 2, 6, 20 minutes, and 1, 2, 4, 8, 12, and 24 hourspost-dose for determining plasma concentrations used to calculate PKparameters. The ratios of clinical spirometry outcomes (FEV1) toglycopyrrolate PK outcomes (AUC0-12 and C_(max)) were determined.

All doses of Glycopyrrolate MDI were safe and well tolerated, and themean plasma glycopyrrolate concentration-time profiles were wellcharacterized with rapidly occurring peak plasma concentrations,generally within 20 minutes. Plasma glycopyrrolate increased with doselevel. FIG. 5 shows the serum glycopyrrolate concentration (in pg/mL)compared to placebo over a 24 hour period experienced in the studysubjects.

Glycopyrrolate MDI showed statistically significant and clinicallyrelevant superior efficacy compared to Placebo MDI (p<0.001 for all fourglycopyrrolate doses) with a clear dose response relationship. Theefficacy of Glycopyrrolate MDI 144 μg and Glycopyrrolate 72 μg bracketedthat of tiotropium 18 μg in terms of peak improvement in FEV₁ over time.For improvement in secondary FEV₁ endpoints relative to test daybaseline, including trough FEV₁ at 12 hours, FEV₁ AUC₀₋₁₂, FEV₁ AUC₀₋₂₄,FEV₁ AUC₁₂₋₂₄, and 12 and 24-hour trough FEV₁, all doses ofGlycopyrrolate MDI demonstrated clinically relevant and statisticalsuperiority compared to Placebo MDI (p≦0.049 for all four dose levels),with the exception of improvement in trough FEV₁ at 24 hours followingadministration of Glycopyrrolate MDI 36 μg (difference compared toplacebo=0.073 L, p=0.059). Similar to the clear dose-responserelationship observed for improvement in peak FEV₁, dose ordering acrossall four doses of Glycopyrrolate MDI evaluated was also observed forimprovements in FEV₁ AUC₀₋₁₂, FEV₁ AUC₀₋₂₄, and FEV₁ AUC₁₂₋₂₄.

The Glycopyrrolate MDI 144 μg and 72 μg doses were shown to bestatistically non-inferior to tiotropium 18 μg in terms of peak changein FEV₁, FEV₁ AUC₀₋₁₂, and FEV₁ AUC₀₋₂₄, with the a priori definednon-inferiority bound of 100 mL. The Glycopyrrolate 144 μg dose was alsonon-inferior to tiotropium for 12-hour trough and FEV₁ AUC₁₂₋₂₄.Point-estimates for the majority of the FEV₁ parameters for the 72 and144 μg doses were within ±50 mL compared to tiotropium. In general, thesecondary endpoints (time to onset of effect, peak and trough FEV₁, FVC,PEFR, and peak IC) confirmed the findings of the primary endpoint.Glycopyrrolate MDI demonstrated a more rapid onset of action compared totiotropium 18 μg, with mean time to ≧10% improvement in FEV₁ of 1 houror less for all doses of Glycopyrrolate MDI evaluated, compared toapproximately 3 hours for tiotropium 18 μg.

FIG. 6 plots the mean change in FEV₁ from baseline (in liters)experienced by the study subjects over a period of 24 hours. FIG. 7depicts the change in FEV₁ from baseline (in liters) for patients atdifferent glycopyrrolate dosing levels compared to the results obtainedfor tiotropium. Specifically, FIG. 7 compares the peak change frombaseline over the placebo value for different glycopyrrolateconcentrations and the area under the curve over a 12 hour and 24 hourperiod. FIG. 8 depicts the proportion of patients which experiencedeither 1) an increase from baseline in FEV₁ of at least 200 mL or 2) a12%, or greater, increase from baseline in FEV₁ coupled with totalincrease in FEV₁ of at least 150 mL or greater. FIG. 9 shows the peakimprovement in IC experienced by patients administered the various dosesof Glycopyrrolate, as well as the peak improvement in IC for patientsreceiving tiotropium. FIG. 10 shows change in FEV₁ cumulatively over a24 hour period in patients receiving glycopyrrolate, compared with theresults obtained from another clinical study where NVA237 (a powderformulation of glycopyrrolate) was given at various doses by Singh et al(D Singh, P A Corris, and S D Snape. “NVA237, a once-daily inhaledantimuscarinic, provides 24-hour bronchodilator efficacy in patientswith moderate to-severe COPD” Poster presented at the American ThoracicSociety International Conference, San Diego, Calif., May 19-24, 2006).

Example 7

Glycopyrrolate (GP) was micronized using a jet mill to a volume medianoptical diameter (d₅₀) of 1.4 μm with 90% of the cumulative distribution(d₉₀) having a volume optical diameter below 3.0 μm. Suspendingparticles were manufactured similarly to those in Example 1. MDIcanisters were manufactured using FEP coated Presspart cans to provideproducts with metered dose of 5.5 μg/actuation GP and 44 μg/actuation GPwhich correlates to approximately 4.5 μg/actuation and 36 μg/actuationGP delivered dose from a 50 μl volume metering chamber from commerciallyavailable Bespak valves. The formulations contained 6 mg/mL ofsuspending particles. The MDI canisters were manufactured using standardpressure filling process where drug substance and the suspending weremixed with HFA 134a in a suspension vessel and filled into canistersthrough a commercially available filling head.

Each lot was tested for delivered dose uniformity through can life andaerodynamic particle size distribution by Next Generation Impactor aftermanufacture. The aerodynamic particle size distributions as measured bythe NGI are shown in FIGS. 11 and 12. Also shown are the amounts of drugrecovered from valve stem and actuator, and from the induction port(throat) and its mouth piece adaptor. Recovered masses are expressed aspercent of nominal dose. The fine particle fraction remained unchangedover 168 cycles, illustrating the stability of the GP co-suspensionsdisclosed herein across a GP dose range. The delivered dose through lifeof the MDI canisters is shown in FIGS. 13 and 14. No change in delivereddose from beginning to middle of can is observed and a ˜10% increasefrom middle to end of canister. The change from middle to end isanticipated based upon evaporative losses of propellant as the can isemptied. The compositions described in this example demonstratedesirable delivered dose uniformity for MDI for doses as low as 4.5μg/actuation.

In addition, canisters from each lot were subjected to a temperaturecycling stability study. The canisters were subjected to −5° C. and 40°C. alternating between temperatures every 6 hours for a total durationof 84 cycles (3 weeks) and 168 cycles (6 weeks). After 184 cycles, the %FPF (ex-actuator) is not significantly different from initial. A summaryof the stability of the fine particle fraction is shown in Table 6.

TABLE 6 Temperature Cycling Stability of the Fine Particle Fraction ofcrystalline GP co suspended with suspending particles at two doses inMDI containing HFA 134a 4.5 μg/actuation 36 μg/actuation Timepoint (%FPF ex-actuator) (% FPF ex-actuator) Initial 60.9 57.4 3 Weeks (84cycles) 61.9 58.0 6 Weeks (168 cycles) 60.6 59.0

Example 8

MDI Canisters were manufactured to contain 6 mg/mL suspending particleconcentration and to provide a metered dose of 36 μg/actuation with a 50μl valve volume according to Example 7. Micronized GP had a d₅₀ and d₉₀of 1.6 μm and 4.1 μm respectively and suspending particles weremanufactured similarly to the process described in Example 1. Thecanisters were placed on stability without protective packaging at 25°C./60% RH and stored for duration of 12 months. Aerodynamic particlesize distribution was determined by next generation impaction at 2weeks, 1, 2, 3, 6 or 12 months. The fine particle fraction, as apercentage of GP ex-actuator, at initial sampling was 50.2%. Nosignificant change in the fine particle fraction was noted at any of thetimepoints out to 12 months, with FPF of 47.7% after 12 months. FIG. 15provides a view of the entire aerodynamic size distribution for each ofthe timepoints demonstrating desirable consistency on aerosol delivery.A summary of the fine particle fraction is shown in Table 7.

TABLE 7 Stability of the Fine Particle Fraction of crystalline GP cosuspended with suspending particles in MDI containing HFA 34a at 25° C.and 60% RH with no protective packaging % FPF Time-Point (ex actuator)Initial 50.2 2 Week 46.1 1 Month 42.0 2 Month 46.0 3 Month 48.9 6 Month47.7 12 Month 47.7

Example 9

MDI Canisters were manufactured to contain 6 mg/mL suspending particleconcentration and to provide a metered dose of 36 μg/actuation asdescribed in Example 7. These canisters were packaged in a heat sealedaluminum foil overwrap containing desiccant, and cycled for 6 weeks (6hours at −5° C. and 6 hours at 40° C.). The delivered dose uniformitythrough use was tested at the 0, 2, 4 and 6 weeks time points. The meanglycopyrrolate delivered dose of each lot each time period was within±15% of the mean, with one exception, as demonstrated in FIG. 16. Theaerodynamic particle size distribution as measured by NGI remainunchanged after 168 temperature cycles as shown in FIG. 17.

Example 10

MDI Canisters were manufactured to contain 6 mg/mL suspending particleconcentration and to provide a metered dose of 24 μg per actuationaccording to Example 7. These canisters were stored for six weeks at 50°C. under ambient humidity. Another lot was stored for 8 weeks at 40° C.and 75% relative humidity. Yet another lot was stored for 12 weeks at40° C. and 75% relative humidity. The fine particle fraction was 59.3%initially. The canister stored for 6 weeks at 50° C. had an FPF that wasunchanged compared to the initial lot, i.e. at 58.4%. The lot stored at40° C. for 8 and 12 weeks had an FPF that was also unchanged compared tothe initial, i.e. at 56.8% and 57.6% respectively. The aerodynamicparticle size distributions as measured by the NGI are shown in FIG. 18.The MMAD remains relatively unchanged after 6 weeks at 50° C., 3.94 μm,and up to 12 weeks at 40° C., 3.84 μm, compared to the initial at 3.54μm. In addition, the FPF and the amounts of glycopyrrolate recoveredfrom valve stem and actuator, and from the induction port (throat) andits mouth piece adaptor, remained relatively unchanged over 3 months atelevated temperatures.

Example 11

Metered dose inhalers including pharmaceutical compositions offormoterol fumarate as described herein were prepared. Formoterolfumarate,(±)-2-hydroxy-5-[(1RS)-1-hydroxy-2-[[(1RS)-2-(4-methoxyphenyl)-1-methylethyl]-amino]ethyl]formanilide fumarate, also known as(±)-2′-hydroxy-5′-[(RS)-1-hydroxy-2-[[(RS)-p-methoxy-α-methylphenethyl]-amine]ethyl]formanilide fumarate, dihydrate was micronized to form active agentparticles. The particle size distribution of the micronized formoterolfumarate (FF) was determined by laser diffraction. 50% by volume of themicronized particles exhibited an optical diameter smaller than 1.6 μm,and 90% by volume exhibited an optical diameter smaller than 3.9 μm.

Suspending particles were manufactured as follows: 503 mL of afluorocarbon-in-water emulsion of PFOB (perfluorooctyl bromide)stabilized by a phospholipid was prepared. 20.6 g of the phospholipid,DSPC (1,2-disteroyl-sn-glycero-3-phosphocholine), and 1.9 g of calciumchloride were homogenized in 403 mL of hot water (75° C.) using a highshear mixer. 100 mL of PFOB were added slowly during homogenization. Theresulting coarse emulsion was then further homogenized using a highpressure homogenizer (Model C3, Avestin, Ottawa, Calif.) at pressures ofup to 170 MPa for 5 passes.

The emulsion was spray dried in nitrogen using the following spraydrying conditions: Inlet temperature 95° C., outlet temperature 71° C.,emulsion feed rate 2.4 mL/min, total gas flow 498 L/min. The particlesize distribution of the suspending particles was determined by laserdiffraction. 50% by volume of the suspending particles were smaller than3 μm, the geometric standard deviation of the distribution was 1.9.

Metered dose inhalers were prepared by weighing the target masses ofmicronized active agent particles and suspending particles into coatedglass vials with 15 mL volume. The target masses and the targetdelivered dose assuming 20% actuator deposition are given in Table 8 forthree different configurations. For each configuration, additional glassbottles were filled with the respective amount of FF active agentparticles without any suspending particles. The canisters were crimpsealed with 63 μl valves (Valois, Les Vaudreuil, France) and filled with11 g (9.1 mL at 25° C.) of HFA 134a (1,1,1,2-tetrafluoroethane) (IneosFluor, Lyndhurst, UK) by overpressure through the valve stem. Afterinjecting the propellant, the canisters were sonicated for 15 secondsand agitated on a wrist action shaker for 30 minutes.

TABLE 8 Target doses for formoterol fumarate co-suspensions of Example10 FF Active Target Suspending Agent Suspending delivered Particle toParticles Particles dose active particle Configuration # μg/can mg/canμg ratio 6A 300 50 1.7 167 6B 860 4.6 58 6C 3010 16.5 16.6

Visual observation of the co-suspended configurations (6A, 6B, 6C)showed no sedimentation of the crystalline FF forming the active agentparticles. The suspension flocculated slowly and formed a homogeneous,single cream layer. For all concentrations tested the micronized activeagent particles alone sedimented quickly. Pictures of the co-suspensionand the traditional comparator suspensions, indicated by an asterisk,are shown in FIG. 19. The vials were left to settle for 24 h withoutagitation. No FF crystals were visible at the bottom of any of theco-suspension vials.

The results showed that the FF crystals associated with the suspendingparticles. The association between FF particles and suspending particleswas strong enough to overcome buoyancy forces, as FF particles did notseparate from the suspending particles and settling of the active agentparticles was successfully inhibited in each of the three differentformulation configurations.

Example 12

Formoterol fumarate MDI compositions were prepared according to thepresent invention. Micronized formoterol fumarate was commerciallyobtained and its particle size distribution measured as described inExample 1 was characterized by a d₁₀, d₅₀, d₉₀ of 0.6, 1.9 and 4.4 μmrespectively and a Span of 2.0. Suspending particles used were preparedin a similar manner described in Example 1. MDI manufacturing wasaccomplished using a drug addition vessel (DVA) by first adding half ofsuspending particle quantity, next filling the microcrystalline FF, andlastly adding the remaining half of suspending particles to the top.Materials were added to the DAV in a humidity controlled environment of<10% RH. The DAV was then connected to a 4 L suspension vessel. A slurrywas then formed by adding a known amount of HFA-134a propellant (IneosFluor, Lyndhurst, UK) into the DAV, which is then removed from thesuspension vessel and gently swirled. The slurry is then transferredback to the suspension mixing vessel and diluted with additionalHFA-134a to form the final suspension at target concentration stirringgently with an impeller. The temperature inside the vessel wasmaintained at 21-23° C. throughout the entire batch production. Afterrecirculation of the batch for 30 min, 14-mL fluorinated ethylenepolymer (FEP) coated aluminum canisters (Presspart, Blackburn, UK) werefilled with the suspension mixture through 50 μL EPDM valves (Bespak,King's Lynn, UK). Sample canisters were then selected at random fortotal canister assay to ensure correct formulation quantities.

The freshly manufactured co-suspension MDI batch was then placed on oneweek quarantine before initial performance analysis. Aerosol performancewas assessed in accordance with USP <601> (United States Pharmacopeiamonograph 601). A Next Generation Impactor (NGI) operated at a flow rateof 30 L/min was used for determination of particle size distribution.Sample canisters were seated into an actuator with two waste actuationsand two additional waste priming actuations. Five actuations werecollected in the NGI with a USP throat attached. The valve, actuator,throat, NGI cups, stages, and filter were rinsed with volumetricallydispensed solvent. The sample solutions were assayed using a drugspecific chromatographic method. The fine particle fraction was definedusing the sum of stages 3 through filter. Delivered dose uniformitythrough use testing was performed using a Dose Uniformity SamplingApparatus as described by USP <601>. Two actuations were collected andassayed at beginning, middle and end of use.

FIG. 20 shows the delivered dose uniformity for a co-suspension of FF ata 4.8 μg target dose per actuation. The individual delivered dose peractuation for beginning, middle and end of actuations was within ±25% ofthe mean delivered dose.

Example 13

Formoterol Fumarate MDI compositions were prepared according to thepresent invention. Micronized formoterol fumarate was commerciallyobtained and its particle size distribution measured as described inExample 1 was characterized by a d₁₀, d₅₀, d₉₀ of 0.6, 1.9 and 4.4 μmrespectively and a Span of 2.0. Suspending particles used were preparedin a similar manner described in Example 1. MDI manufacturing wasaccomplished as described in Example 12.

Aerosol performance was assessed in accordance with USP <601>. A NextGeneration Impactor (NGI) operated at a flow rate of 30 L/min was usedfor determination of particle size distribution. Sample canisters wereseated into an actuator with two waste actuations and two additionalwaste priming actuations. Five actuations were collected in the NGI witha USP throat attached. The valve, actuator, throat, NGI cups, stages,and filter were rinsed with volumetrically dispensed solvent. The samplesolutions were assayed using a drug specific chromatographic method. Thefine particle fraction was defined using the sum of stages 3 throughfilter. The aerodynamic particle size distribution of a FF co-suspensionformulation was evaluated after manufacture and after three months ofstorage at 25° C. and 75% RH (unprotected canisters) and 40° C. and 75%RH (protected canisters wrapped in aluminum foil pouch). The aerodynamicparticle size distributions shown in FIG. 21 demonstrate that thecompositions described in the present invention display desirablestability characteristics even at accelerated conditions.

Example 14

The chemical stability of formoterol fumarate (FF) included in aco-suspension formulation prepared according Example 11 was evaluated.FF MDI canisters containing HFA 134a were overwrapped with an aluminumfoil pouch and stored at 25° C. and 60% relative humidity and 40° C. and75% relative humidity for thirteen and six months, respectively.Likewise FF MDI canisters containing HFA 227ea were overwrapped with analuminum foil pouch and stored at 25° C. and 60% relative humidity and40° C. and 75% relative humidity for six months. The amount of impurityF, a characteristic degradation product of FF, and total impurities weredetermined by reverse phase HPLC assay as follows: each canister ischilled, cut open, and the can contents are transferred to a centrifugetube; the contents were dissolved in organic solvent, followed by theaddition of an aqueous solvent to precipitate excipient (DSPC) from thesolution; the solution was centrifuged to produce a clear supernatantsolution; and each sample solution was analyzed using a C18 column,4.6×150 mm and 3.0 μm particle size. The column temperature was kept at30° C. The injection volume was 20 μl, and flow rate was set at 1 mL/minand detected by determining the UV absorption at 214 nm. A gradient wasused mixing pH 3.1 aqueous phosphate buffer and acetonitrile, 17%acetonitrile first 27 minutes, then 50% acetonitrile for 30 secondsfollowed by 6.5 minutes at 75% acetonitrile and 17% acetonitrile for 8minutes. Impurities were reported as area percent of formoterol peakarea (corrected for relative response factors, where available). Asshown in FIG. 22 (or Table 9 and 10), a co-suspension prepared usingcrystalline FF active agent particles suspended in HFA 134a withsuspending particles was chemically stable for 18 months at atemperature of 25° C. and 60% relative humidity, in contrast a spraydried, non co-suspended formoterol formulation showed a fasterdegradation rate under the same storage conditions. Likewise crystallineFF active agent particles formed a chemically stable co-suspension inHFA 227a, as shown in Table 11.

TABLE 9 Chemical Stability of Spray Dried FF Suspending Particles in FFMDI Containing HFA 134a at 25° C./60% RH, Overwrapped in Aluminum FoilPouches Time (months) 0 2 3 12 18 Impurity F (%) ND 0.12% 0.04% 1.16%2.77% Total Impurities (%) 0.62% 1.42% 1.75% 2.33% 4.39% ND = Notdetected

TABLE 10 Chemical Stability of Crystalline FF Co-suspended withSuspending Particles in FF MDI Containing HFA 134a at 25° C./60% RH,Overwrapped in Aluminum Foil Pouches Time (months) 0 1 2 3 6 10 13Impurity F (%) 0.05% 0.08% 0.08% 0.14% 0.06% 0.22% 0.35% TotalImpurities (%) 0.44% 0.32% 0.32% 0.37% 0.18% 0.45% 0.64% at 40° C./75%RH, Overwrapped in Aluminum Foil Pouches Time (months) 0 1 2 3 6Impurity F (%) 0.05% 0.11% 0.31% 1.18% 1.74% Total Impurities (%) 0.44%0.41% 0.75% 1.58% 2.54%

TABLE 11 Chemical Stability of Crystalline FF Co-suspended withSuspending Particles in FF MDI Containing HFA 227ea at 25° C./60% RH,Overwrapped in Aluminum Foil Pouches Time (months) 0 1 2 3 6 Impurity F(%) 0.04 0.06 0.07 0.13 0.05 Total Impurities (%) 0.4 0.3 0.3 0.4 0.1 at40° C./75% RH, Overwrapped in aluminum foil pouches Impurity F (%) 0.040.08 0.18 0.80 1.14 Total Impurities (%) 0.40 0.39 0.53 1.13 1.56

Example 15

Micronized formoterol fumarate dihydrate (FF) (Inke, S. A., Barcelona,Spain) used in the present example had with particle size distributionby laser diffraction of 50% by volume of the micronized particlesexhibited an optical diameter smaller than 1.9 μm, 90% by volumeexhibited an optical diameter smaller than 4.1 μm. Four batches ofsuspending particles were manufactured by spray drying as described inExample 1. All four batches were spray-dried from aqueous solution;solution concentration and spray drying parameters are given in Table12.

TABLE 12 Suspending particle configurations used in Example 15 SprayDrying Parameters Feed Total Particle Size C_(f) rate Gas Distributionin in T_(in) T_(out) Flow VMD Powder mg/ mL/ in in in std in #Composition mL min ° C. ° C. L/min μm GSD XA 100% 80 10 150 82 385 1.622.20 trehalose XB 100% HP-β- 80 10 100 68 885 1.61 2.21 cyclodextrin XC100% Ficoll 80 10 100 70 885 1.19 2.27 PM 70 XD 100% Inulin 80 10 100 70885 1.23 2.20

Electron micrographs of the suspending particles showed a variety ofmorphologies, and are shown in FIG. 23 through FIG. 26, with FIG. 23providing a micrograph of trehalose suspending particles, FIG. 24providing a micrograph of HP-β-cyclodextrin suspending particles, FIG.25 providing a micrograph of Ficoll MP 70 suspending particles, and FIG.26 providing a micrograph of inulin suspending particles. Trehaloseparticles appear to be spherical, with a smooth surface.HP-β-cyclodextrin particles show extensive wrinkling of the surface,suggesting a partially buckled exterior with a hollow core. Ficoll MP 70and Inulin particles display some surface rugosity but are generallyspheroidal.

Metered dose inhalers were prepared by weighing 0.9 mg of the micronizedFF active agent particles and 60 mg of suspending particles into coatedglass vials with 15 mL volume. FF was combined with each type of thefour suspending particle species of Table 11. The canisters were crimpsealed with 50 μL valves (Valois DF31/50 RCU, Les Vaudreuil, France) andfilled with 10 mL of HFA propellant 134a (Ineos Fluor, Lyndhurst, UK) byoverpressure through the valve stem. After injecting the propellant, thecanisters were sonicated for 30 seconds and agitated on a wrist actionshaker for 30 minutes. Additional inhalers containing suspendingparticles only and active agent particles only were filled as a controlfor each configuration.

Crystalline FF has a greater density than propellant 134a at roomtemperature, as do all four species of suspending particles in thepresent example. Consequently both FF and suspending particles settledto the bottom of the inhalers at room temperature. To test theseinhalers for active-suspending agent particle interactions indicating aco-suspension, the inhalers were immersed in an ethanol bath at ≦−10° C.(resulting in increased propellant density) and allowed to equilibratefor a minimum of 30 minutes. At this temperature, the FF active agentparticles are less dense than the propellant and consequently cream tothe top of the propellant volume, while all four species of suspendingagent particles remain settled at the bottom of the propellant volume.

The tested configurations and the results of the observations arepresented in Table 13. FF active agent particles alone formed a creamlayer atop the propellant volume, and trehalose, HP-β-cyclodextrin,inulin, and Ficoll PM70 particles alone all settled to the bottom of theglass vial. FF active agent particles in combination with trehalosesuspending particles formed a single sediment layer, with no particlescreamed or afloat in the propellant, indicating that the FF particlesinteract with the trehalose suspending particles, and a co-suspension isformed. In the case of FF particles in combination withHP-β-cyclodextrin suspending particles, some turbidity was present inthe propellant, similar to that observed in the suspending particle onlycontrol vial. Additionally, some floating flocs were observed, which mayhave been FF particles; however, such flocs accounted for a small amountof solid mass relative to the control vial, indicating that some if notall FF particles were interacting with the suspending agent particles.Thus, this configuration is an example of a partial co-suspension. FFparticles in combination with inulin suspending particles formed asingle sediment layer, indicating a co-suspension was formed. Thoughsome turbidity was present in this configuration, similar cloudiness wasobserved in the inulin-only control vial. FF active agent particles incombination with Ficoll PM70 suspending particles formed a sedimentlayer at the bottom of the vial, indicating that a co-suspension wasformed. While some turbidity and floating flocs were observed in thisconfiguration, similar turbidity, and floc frequency were observed inthe Ficoll-only control vial.

TABLE 13 Summary of tested configurations and results of observationsSuspending Particle to Active Container Contents in 10 mL Particle Co-ID p134a Ratio Observational Notes, ≦−10° C. suspension 0-FF 0.9 mg FFn/a Creamed to top n/a T 60 mg trehalose n/a Settled to bottom n/a T-FF60 mg 67 Sediment layer; no particles Yes trehalose, 0.9 mg creamed FF C60 mg HP-β- n/a Settled to bottom; some n/a cyclodextrin turbidity C-FF60 mg HP-β- 67 Solids mostly in sediment layer partial cyclodextrin, 0.9mg at bottom; some turbidity; some FF floating flocs present I 60 mgInulin n/a Settled to bottom; some n/a turbidity I-FF 60 mg Inulin, 67Sediment layer; no particles Yes 0.9 mg FF creamed; some turbidity F 60mg Ficoll n/a Settled to bottom, with some n/a PM70 floating flocs F-FF60 mg Ficoll 67 Sediment layer; very few Yes PM70, 0.9 mg floating flocsFF

Example 16

Co-suspension compositions including glycopyrrolate (GP) and formoterolfumarate (FF) active agent particles were produced and MDIsincorporating the co-suspension compositions were prepared. Theco-suspension compositions produced included GP active agent particles,FF active agent particles or a combination of both GP and FF activeagent particles. The GP and FF material was supplied as micronized,crystalline material with particle size distribution as shown in Table14.

Suspending particles were manufactured via spray dried emulsion at afeed stock concentration of 80 mg/mL with a composition of 93.44% DSPC(1,2-Distearoyl-sn-Glycero-3-Phosphocholine) and 6.56% anhydrous calciumchloride (equivalent to a 2:1 DSPC:CaCl₂ mole/mole ratio). During theemulsion preparation, DSPC and CaCl₂ was dispersed with a high shearmixer at 8000-10000 rpm in a vessel containing heated water (80±3° C.)with PFOB slowly added during the process. The emulsion was thenprocessed with 6 passes in a high pressure homogenizer (10000-25000psi). The emulsion was then spray dried via a spray dryer fitted with a0.42″ atomizer nozzle with a set atomizer gas flow of 18 SCFM. Thedrying gas flow rate was set to 72 SCFM with an inlet temperature of135° C., outlet temperature 70° C., and an emulsion flow rate of 58mL/min.

The co-suspensions were prepared by first dispensing the appropriatequantities of micronized GP and FF active agent particles and suspendingparticles into a drug addition vessel (DAV) inside a humidity controlledchamber (RH<5%). In the present Example, the suspending particles wereadded in three equal portions intercalating the addition of GP and FFafter the first and second addition respectively. The DAV is then sealedunder a nitrogen atmosphere and connected to the suspension vesselcontaining 12 kg of HFA-134a (Ineos Fluor, Lyndhurst, UK). A slurry wasthen formed by adding 0.5-1 kg of HFA-134a into the DAV, which is thenremoved from the suspension vessel and gently swirled. The slurry isthen transferred back to the suspension mixing vessel and diluted withadditional HFA-134a to form the final suspension at target concentrationstirring gently with an impeller. The suspension is then recirculatedvia a pump to the filling system for a minimum time prior to initiationof filling. Mixing and recirculation continue throughout the fillingprocess. 50 μL valves (Bespak, King's Lynn, UK) are placed onto 14-mLfluorinated ethylene polymer (FEP) coated aluminum canisters (Presspart,Blackburn, UK) canisters and then purged of air either by a vacuumcrimping process, or an HFA-134a purging process followed by valvecrimping. The crimped canisters are then filled through-the-valve withthe appropriate quantity of suspension, adjusted by the meteringcylinder.

TABLE 14 Glycopyrrolate and Formoterol Fumarate particle sizedistributions. Designation d₁₀ (μm) d₅₀ (μm) d₉₀ (μm) Span FF API 0.61.9 4.1 1.8 GP API 0.5 1.3 3.0 1.9

MDIs containing the dual co-suspensions described in this Example wereprepared to contain two different doses GP and FF. Specifically, a firstrun of dual co-suspension compositions were prepared to provide 18 μgper actuation GP and 4.8 μg per actuation FF (“low dose”), and a secondrun of dual co-suspension compositions were prepared to provide 36 μgper actuation GP and 4.8 μg per actuation FF (“high dose”). In additionto the dual co-suspensions compositions, co-suspensions including asingle species of active agent particle were prepared. Thesecompositions included either GP active agent particles or FF activeagent particles and were referred to as “mono” or “monotherapy”co-suspensions. The monotherapy co-suspension compositions were preparedas described for the dual co-suspensions, except that they included onlyone species of active agent particles (either GP or FF). The monotherapyco-suspensions were formulated and monotherapy MDIs prepared to providethe following targeted delivered doses: 18 μg per actuation of GP, and0.5, 1.0, 3.6 or 4.8 μg per actuation of FF. The compositions and MDIsproviding 0.5 μg FF and 1 μg FF per actuation are referred to as “ultralow” dose and were manufactured in a similar manner at a 4 L scale.

The drug specific aerodynamic size distributions achieved with MDIscontaining the co-suspension compositions prepared according to thisExample were determined as described in Example 1. The proportionalityof the aerodynamic size distributions of GP obtained from the low andhigh dose dual co-suspensions as well as the equivalency between thedual and monotherapy co-suspensions is demonstrated in FIG. 27. In thesame manner, the proportionality of the aerodynamic size distributionsof FF obtained from the dual and monotherapy co-suspensions, includingthe ultralow, low, and high dose compositions is demonstrated in FIG.28.

The delivered dose uniformity of the ultra low dose FF monotherapy MDIswas also measured as described in Example 1. The DDU for the FF MDIcontaining 0.5 μg per actuation and 1.0 μg per actuation are shown inFIG. 29. Desirable dose delivery uniformity is achieved demonstratingthe utility of the present invention to consistently deliver ultra lowdoses. In order to evaluate whether the combination of GP and FF withina single formulation would result in the degradation of the aerosolproperties relative to compositions including a single active agent, theaerosol properties of co-suspension compositions were assessed relativeto suspension compositions including only a single active agent. As canbe seen in FIG. 30, the aerosol performance of the combinationco-suspension composition including both GP and FF active agent was nodifferent than the aerosol performance achieved by suspensioncompositions including either GP or FF alone. Therefore, there were nocombination effects observed.

Example 17

Micronized salmeterol xinafoate(4-hydroxy-α1-[[[6-(4-phenylbutoxy)hexyl]amino]methyl]-1,3-benzenedimethanol, 1-hydroxy-2-naphthalenecarboxylate) wasreceived by the manufacturer (Inke SA, Germany) and used as active agentparticles. The particle size distribution of the salmeterol xinafoate(SX) was determined by laser diffraction. 50% by volume of themicronized particles exhibited an optical diameter smaller than 2 μm,90% by volume exhibited an optical diameter smaller than 3.9 μm.

Suspending particles were manufactured as follows: 150 mL of afluorocarbon-in water emulsion of PFOB (perfluoroctyl bromide)stabilized by a phospholipid was prepared. 12.3 g of the phospholipid,DSPC (1,2-Distearoyl-sn-Glycero-3-Phosphocholine), and 1.2 g of calciumchloride were homogenized in 100 mL of hot water (70° C.) using a highshear mixer. 65 mL of PFOB were added slowly during homogenization. Theresulting coarse emulsion was then further homogenized using a highpressure homogenizer (Model C3, Avestin, Ottawa, Calif.) at pressures ofup to 140 MPa for 3 passes

The emulsion was spray dried in nitrogen using the following spraydrying conditions: Inlet temperature 90° C., outlet temperature 69° C.,emulsion feed rate 2.4 mL/min, total gas flow 498 l/min. The particlesize distribution of the suspending particles, VMD, was determined bylaser diffraction. 50% by volume of the suspending particles weresmaller than 2.7 μm, the Geometric Standard Deviation of thedistribution was 2.0. Additionally, the aerodynamic particle sizedistribution of the suspending particles was determined with atime-of-flight particle sizer. 50% by volume of the suspending particleshad an aerodynamic particle diameter smaller than 1.6 μm. The largedifference between aerodynamic particle diameter and optical particlediameter indicates that the suspending particles had a low particledensity <0.5 kg/L.

Metered dose inhalers were prepared by weighing 2 mg of SX active agentparticles and 60 mg of suspending particles into fluorinated ethylenepolymer (FEP) coated aluminum canisters (Presspart, Blackburn, UK) with19 mL volume. The suspending particle to active particle ratio was 30.The target delivered dose assuming 20% actuator deposition was 10 μg.The canisters were crimp sealed with 63 μl valves (# BK 357, Bespak,King's Lynn, UK) and filled with 10 mL of HFA 134a(1,1,1,2-tetrafluoroethane) by overpressure through the valve stem.After injecting the propellant, the canisters were sonicated for 15seconds and agitated on a wrist action shaker for 30 minutes. Thecanisters were fitted with polypropylene actuators with a 0.3 mm orifice(# BK 636, Bespak, King's Lynn, UK). Additional inhalers for visualobservation of suspension quality were prepared using 15 mL glass vialsincluding a comparator filled with micronized SX only. Aerosolperformance was assessed as described in Example 1. The MMAD was 3.7 μmand the fine particle fraction was 48%. Because the SX crystals formingthe active agent particles and the propellant were nearly densitymatched at 15° C.-20° C., the visual observation was conducted on glassvials that were heated up to 30° C.-35° C. in a water bath. Under theseconditions the SX active agent particles formulated alone sedimentedrapidly, but no SX crystals were visible at the bottom of theco-suspension vial.

Micronized salmeterol xinafoate active agent particles were co-suspendedthrough association with suspending particles of low density that wereformulated according to the disclosure provided herein. The associationbetween salmeterol crystals and the suspending particles was strongenough to overcome buoyancy forces as it was observed that settling ofthe crystals is inhibited.

1.-92. (canceled)
 93. A pharmaceutical composition deliverable from a metered dose inhaler, comprising: a suspension medium comprising a pharmaceutically acceptable propellant; a plurality of active agent particles comprising an active agent selected from a long-acting muscarinic antagonist (LAMA) active agent and a long-acting β₂ adrenergic receptor agonist (LABA) active agent; and a plurality of respirable suspending particles, wherein the respirable suspending particles are substantially insoluble in the suspension medium and comprise a phospholipid; and wherein the ratio of the total mass of the respirable suspending particles to the total mass of the active agent particles is from greater than 1:1 up to 200:1.
 94. The pharmaceutical composition according to claim 93, wherein the suspending particles comprise perforated microstructures.
 95. The pharmaceutical composition according to claim 94, wherein the perforated microstructures are prepared using a spray drying process.
 96. The pharmaceutical composition according to claim 95, wherein the perforated microstructures comprise a spray dried emulsion of perfluorooctyl bromide, DSPC, and calcium chloride in water.
 97. The pharmaceutical composition according to claim 93, wherein the suspending particles exhibit an MMAD selected from between about 10 μm and about 500 nm, between about 5 μm and about 750 nm, between about and 1 μm and about 3 μm.
 98. The pharmaceutical composition according to claim 93, wherein the suspending particles exhibit a volume median optical diameter selected from between about 0.2 μm and about 50 μm, between about 0.5 μm and about 15 μm, between about 1.5 μm and about 10 μm, and between about 2 μm and about 5 μm.
 99. The pharmaceutical composition according to claim 93, wherein the propellant comprises a propellant selected from an HFA propellant, a PFC propellant and combinations thereof, and wherein the propellant is substantially free of additional constituents.
 100. The pharmaceutical composition according to claim 93, wherein a ratio of the total mass of the suspending particles to the total mass of the active agent particles is selected from above about 1.5, up to about 5, up to about 10, up to about 15, up to about 17, up to about 20, up to about 30, up to about 40, up to about 50, up to about 60, up to about 75, up to about 100, up to about 150, and up to about
 200. 101. The pharmaceutical composition according to claim 93, wherein the active agent particles comprise crystalline active agent.
 102. The pharmaceutical composition according to claim 93, wherein the active agent particles comprise micronized, crystalline active agent.
 103. The pharmaceutical composition according to claim 93, wherein the active agent included in the active agent particles is a LAMA active agent selected from glycopyrrolate, dexpirronium, tiotropium, trospium, aclidinium, and darotropium, and any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 104. The pharmaceutical composition according to claim 103, wherein the active agent particles comprise glycopyrrolate, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 105. The pharmaceutical composition according to claim 104, wherein the active agent particles comprise crystalline glycopyrrolate, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 106. The pharmaceutical composition according to claim 104, wherein the active agent particles comprise micronized, crystalline glycopyrrolate, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 107. The pharmaceutical composition according to claim 104, wherein the glycopyrrolate active agent particles are included in the suspension medium at a concentration sufficient to provide a delivered dose of glycopyrrolate per actuation of the metered dose inhaler of no more than 10 μg.
 108. The pharmaceutical composition of claim 107, wherein the glycopyrrolate active agent particles comprise a pharmaceutically acceptable salt of glycopyrrolate selected from fluoride, chloride, bromide, iodide, nitrate, sulfate, phosphate, formate, acetate, trifluoroacetate, propionate, butyrate, lactate, citrate, tartrate, malate, maleate, succinate, benzoate, p-chlorobenzoate, diphenyl-acetate or triphenylacetate, o-hydroxybenzoate, p-hydroxybenzoate, 1-hydroxynaphthalene-2-carboxylate, 3-hydroxynaphthalene-2-carboxylate, methanesulfonate, and benzenesulfonate salts.
 109. The pharmaceutical composition of claim 108, wherein the pharmaceutically acceptable salt of glycopyrrolate is selected from fluoride, chloride, bromide, and iodide salts.
 110. The pharmaceutical composition of claim 109, wherein the pharmaceutically acceptable salt of glycopyrrolate is 3-[(cyclopentyl-hydroxyphenylacetyl)oxy]-1,1-dimethylpyrrolidinium bromide.
 111. A pharmaceutical composition according to claim 93, wherein the active agent included in the active agent particles is a LABA active agent selected from bambuterol, clenbuterol, formoterol, salmeterol, carmoterol, milveterol, indacaterol, and saligenin- or indole-containing and adamantyl-derived β2 agonists, and any pharmaceutically acceptable salts, esters, isomers or solvates thereof.
 112. The pharmaceutical composition according to claim 111, wherein the active agent particles comprise formoterol, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 113. The pharmaceutical composition according to claim 111, wherein the active agent particles comprise crystalline formoterol, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 114. The pharmaceutical composition according to claim 111, wherein the active agent particles comprise micronized, crystalline formoterol, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 115. The pharmaceutical composition according to claim 111, wherein the formoterol active agent particles are included in the suspension medium at a concentration sufficient to provide a delivered dose of formoterol per actuation of the metered dose inhaler of no more than 5 μg.
 116. The pharmaceutical composition according to claim 115, wherein the formoterol active agent comprise a pharmaceutically acceptable salt of formoterol selected from hydrochloric, hydrobromic, sulfuric, phosphoric, fumaric, maleic, acetic, lactic, citric, tartaric, ascorbic, succinic, glutaric, gluconic, tricarballylic, oleic, benzoic, p-methoxybenzoic, salicylic, o- and p-hydroxybenzoic, p-chlorobenzoic, methanesulfonic, p-toluenesulfonic, and 3-hydroxy-2-naphthalene carboxylic acid salts.
 117. The pharmaceutical composition according to claim 116, wherein the pharmaceutically acceptable salt of formoterol is formoterol fumarate.
 118. A method for treating a pulmonary disease or disorder in a patient, the method comprising administering a therapeutically effective amount of a pharmaceutical composition from a metered dose inhaler, the pharmaceutical composition comprising: a suspension medium comprising a pharmaceutically acceptable propellant; a plurality of active agent particles comprising an active agent selected from a LAMA active agent and a LABA active agent; and a plurality of respirable suspending particles, wherein the respirable suspending particles are substantially insoluble in the suspension medium and comprise a phospholipid; and wherein the ratio of the total mass of the respirable suspending particles to the total mass of the active agent particles is from greater than 1:1 up to 200:1.
 119. The method of claim 118, wherein the active agent particles comprise crystalline active agent.
 120. The method of claim 118, wherein the active agent particles comprise micronized, crystalline active agent.
 121. The method of claim 118, wherein the active agent comprises a LAMA active agent selected from glycopyrrolate, dexpirronium, tiotropium, trospium, aclidinium, and darotropium, and any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 122. The method of claim 118, wherein the active agent comprises a LABA active agent selected from bambuterol, clenbuterol, formoterol, salmeterol, carmoterol, milveterol, indacaterol, and saligenin- or indole-containing and adamantyl-derived β₂ agonists, and any pharmaceutically acceptable salts, esters, isomers, or solvates thereof.
 123. The method of claim 118, wherein the pulmonary disease or disorder is selected from at least one of asthma, COPD, allergic rhinitis, sinusitis, pulmonary vasoconstriction, inflammation, allergies, impeded respiration, respiratory distress syndrome, pulmonary hypertension, pulmonary inflammation associated with cystic fibrosis, and pulmonary obstruction associated with cystic fibrosis.
 124. The method of claim 121, wherein said administering the pharmaceutical composition comprises administering a delivered dose of glycopyrrolate, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof, of no more than 150 μg.
 125. The method of claim 124, wherein the glycopyrrolate or pharmaceutically acceptable salt, ester, isomer, or solvate thereof, comprises crystalline glycopyrrolate.
 126. The method of claim 124, wherein the glycopyrrolate or pharmaceutically acceptable salt, ester, isomer, or solvate thereof, comprises micronized, crystalline glycopyrrolate.
 127. The pharmaceutical composition of claim 124, wherein the glycopyrrolate active agent particles comprise a pharmaceutically acceptable salt of glycopyrrolate and the pharmaceutically acceptable salt of glycopyrrolate is 3-[(cyclopentyl-hydroxyphenylacetyl)oxy]-1,1-dimethylpyrrolidinium bromide.
 128. The method according to claim 124, wherein said administering results in a clinically significant increase in inspiratory capacity (IC) in the patient.
 129. The method of claim 121, wherein said administering the pharmaceutical composition comprises administering a delivered dose of glycopyrrolate, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof, of no more than 80 μg, and said administering results in an increase in FEV₁ of at least 150 mL within 0.5 hours, or less.
 130. The method according to claim 122, wherein said administering of the pharmaceutical composition comprises delivering a dose of 10 μg, or less, of formoterol, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof, per actuation of the metered dose inhaler.
 131. The method of claim 130, wherein the formoterol, or pharmaceutically acceptable salt, ester, isomer, or solvate thereof, comprises crystalline formoterol.
 132. The method of claim 130, wherein the formoterol, or pharmaceutically acceptable salt, ester, isomer, or solvate thereof, comprises micronized, crystalline formoterol.
 133. The pharmaceutical composition of claim 130, wherein the active agent particles comprise a pharmaceutically acceptable salt of formoterol and the pharmaceutically acceptable salt of formoterol is formoterol fumarate.
 134. The method according to claim 122, wherein said administering of the pharmaceutical composition comprises delivering a dose of 10 μg, or less, of formoterol, including any pharmaceutically acceptable salts, esters, isomers, or solvates thereof, per actuation of the metered dose inhaler, and said administering of the pharmaceutical composition results in a clinically significant increase in FEV₁ in the patient. 